Method for preparing a liquid composition containing Compound I and use thereof in myocardial perfusion PET imaging

Abstract

The present application provides a method for preparing a liquid composition containing Compound I, which comprises a step of purifying a crude product containing Compound I by high performance liquid chromatography, and its use in myocardial perfusion PET imaging, wherein the mobile phase used in the purification step by high performance liquid chromatography contains ethanol and water. By optimizing the process parameters and process flow, it can be applied to large-scale batch activity production and meet the needs of automation. The resulting myocardial perfusion PET contrast agent has high radiochemical purity, high activity concentration, good product stability, and high and stable yield or output.

Description

Technical Field

[0001] This application belongs to the field of chemical pharmaceutical technology, and particularly relates to a method for preparing a liquid composition containing compound I and its use in myocardial perfusion PET imaging.

Background Art

[0002] Myocardial perfusion imaging began to be used in the non-invasive examination of heart diseases in the 1970s, and its great diagnostic value has been widely accepted worldwide. It has become one of the most important imaging methods in the diagnosis, evaluation of treatment effectiveness, and prognostic diagnosis of coronary heart disease. Myocardial perfusion single photon emission computed tomography (SPECT) imaging technology is the main non-invasive perfusion imaging method currently used in clinical detection of coronary heart disease. However, compared with SPECT, positron emission tomography (PET) imaging has higher spatial and temporal resolutions, can effectively reduce tissue attenuation, and can achieve absolute quantification of coronary blood flow using standard tissue attenuation correction methods. Furthermore, the short half-life of positron radionuclides can effectively reduce the radiation dose around the target tissue, and the short half-life can also shorten the interval between resting imaging and exercise imaging. Commonly used myocardial perfusion PET contrast agents include 15 O-H2O, 13 N-NH3·H2O, 82 Rb, etc. However, the half-lives of the above contrast agents are all short, and their clinical applications are still greatly limited. Compared with other positron radionuclides, 18 F has a long half-life (t 1 / 2 = 109.8 minutes), low positron energy (average energy 249.8 keV), less radiation damage to normal tissues, and its van der Waals radius (1.35) is similar to that of hydrogen (1.2), and it does not affect the biological activity of the labeled compound. Therefore, the development of new fluorine-18 labeled myocardial perfusion contrast agents has important practical significance.

[0003] Compound I has the chemical name 2-tert-butyl-4-chloro-5-((3-((4-((2-(2-fluoro 18 F]ethoxy)ethoxy)methyl))-1H-1,2,3-triazol-1-yl)methyl)benzyl)oxy)pyridazin-3(2H)-one, contains the radionuclide 18 F, can be used in positron emission tomography (PET) imaging, has the characteristic that the content of the chemical substance is very small and a pure product cannot be isolated, and most of the prepared products exist in a solution state.

Disclosure of the Invention

[0004] In the prior art, in the preparation of Compound I, there is a problem that the concentration of 18 F ions added is high, but the yield of the final product is low. Furthermore, the radiochemical purity of the prepared Compound I is not high, especially when left for a certain period of time, the radiochemical purity decreases significantly, and the stability cannot meet the application requirements.

[0005] The object of the present application is to provide a large batch of liquid compositions of Compound I that meet the quality requirements such as high radiochemical purity, high activity concentration, good product stability, high and stable yield or yield.

[0006] The object of the present application is to provide an automatic preparation method for a large batch of liquid compositions of Compound I to meet the needs of clinical pharmaceuticals and design a continuous and stable production process. The technical route provided by the present application can meet the needs of a large amount of initial 18 F activity labeling, optimize the usage amount of the reaction precursor, reaction time, and purification HPLC conditions, improve the reaction yield, and improve the radiochemical purity and stability of Compound I.

[0007] The technical solution of the present application is as follows. 1. A method for preparing a liquid composition of Compound I, comprising the following steps: comprising the step of purifying the crude product containing the compound I by high performance liquid chromatography, the mobile phase used in the purification step by high performance liquid chromatography contains ethanol and water, the compound I is 2-tert-butyl-4-chloro-5-((3-((4-((2-(2-fluoro 18 F]ethoxy)ethoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)benzyl)oxy)pyridazin-3(2H)-one, the preparation method, wherein the mobile phase further contains one or more of sodium vitamin C, vitamin C, and gentisic acid. 2. In the purification step by high performance liquid chromatography, in the mobile phase, the ethanol is 0.2 to 2 parts by volume with respect to 1 part by volume of water, the preparation method according to item 1. 3. In the mobile phase, the ethanol is 0.4 to 1 part by volume with respect to 1 part by volume of water, the preparation method according to item 2. 4. In the purification step by high performance liquid chromatography, the addition amount of the sodium vitamin C is 0 to 20 mg / mL, preferably 0.2 to 10 mg / mL, the preparation method according to item 1. 5. In the purification step by high performance liquid chromatography, the addition amount of the vitamin C is 0 to 10 mg / mL, preferably 0.1 to 5 mg / mL, the preparation method according to item 1. 6. In the purification step by high performance liquid chromatography, the addition amount of the gentisic acid is 0 to 10 mg / mL, preferably 0.1 to 5 mg / mL, the preparation method according to item 1. 7. The chromatography column used in the purification step by the high-performance liquid chromatography is a silica gel column, preferably a reverse-phase C18 silica gel column, more preferably an XBridge BEH C18 OBD Prep column, preferably, isocratic elution is used for purification, and the elution flow rate of the mobile phase is 3 to 6 mL / min. The preparation method according to item 1. 8. Before the purification step by the high-performance liquid chromatography, it further includes a step of performing a nucleophilic substitution reaction. In the nucleophilic substitution reaction, the activated 18 F ions are mixed with the precursor-containing solution of Compound I to perform a nucleophilic substitution reaction to produce a crude product containing Compound I. The name of the precursor of Compound I is methyl 2-(2-((1-(3-(((1-(tert-butyl)))-5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)oxy)methyl)benzyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethyl-4-methylbenzenesulfonate. The preparation method according to item 1. 9. In the nucleophilic substitution reaction, the reaction solvent is an aprotic polar solvent, and the range of the ratio of the dosage of the precursor of Compound I / 18 the activity of F ions is (0.5 to 8):1, the reaction temperature is 90 to 140 °C, the reaction time is 5 to 60 minutes, and it is a sealed reaction. The unit of the dosage of the precursor of Compound I is mg. 18 The unit of the activity of F ions is Ci. The preparation method according to item 8. 10. The aprotic polar solvent is one or more of acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, and 2-methyl-2-butanol. The preparation method according to item 9. 11. Before the step of the nucleophilic substitution reaction, 18 it further includes a preparation step of F ions. The 18 preparation step of F ions is 18Preparation of F-ion solution, 18 Concentration and elution of F-ions, and 18 Further includes activation of F-ions, More preferably, The 18 In the preparation of the F-ion solution, an accelerator is used to 18 Prepare the F-ion solution, The 18 In the concentration of the F-ions, the prepared 18 F-ion solution is concentrated through an anion exchange cartridge, The 18 In the elution of the F-ions, a cryptand and an alkali metal salt catalyst solution are used to 18 Elute the F-ions, The 18 In the activation of the F-ions, the temperature is programmed to dry the solvent with nitrogen or other inert gas, 18 Activate the F-ions and obtain the activated 18 F-ions, The preparation method according to item 8. 12. In the step of preparing the 18 F-ion solution, 18 Water containing O is transferred to the target position of the accelerator, and the accelerator is started to generate a proton beam to 18 Collide with the water containing O, 18 Generate a solution containing F-ions, and the 18 Initial activity of F is 0.045 Ci to 11 Ci, preferably, the 18 Initial activity of the F-ions is 3.15 Ci to 11 Ci, The preparation method according to item 11. 13. In the step of concentrating the 18 F-ions, The anion exchange cartridge is a tetraalkylammonium salt anion exchange cartridge, The preparation method according to item 11. 14. In the step of eluting the 18 F-ions, In the catalyst solution of the cryptand and the alkali metal salt, the dosage of the cryptand is 5 to 40 mg, and the dosage of the alkali metal salt is 1.5 to 20 mg. Preferably, the cryptand is 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8,8,8]hexacosane (cryptofix-2.2.2, aminopolyether), and the alkali metal salt is one or more of K2CO3, Na2CO3, Cs2CO3, KHCO3, and NaHCO3. More preferably, the catalyst solution is selected from a mixed solvent system of acetonitrile and water, and the volume ratio of acetonitrile to water is (0.2 to 10):1. The preparation method according to item 11. 15. In the 18 In the activation step of F ions, the activation temperature is 80 to 130 °C. The control according to the temperature program includes the following steps: 100 to 120 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation 60 to 120 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation 150 to 200 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -60 to -100 mbar, evaporation 10 to 30 seconds; 100 to 120 °C, positive pressure 800 to 1200 mbar, vacuum pressure -800 to -1000 mbar, evaporation 80 to 120 seconds; 80 to 100 °C, positive pressure 400 to 600 mbar, vacuum pressure -800 to -1000 mbar, evaporation 100 to 120 seconds; 80 to 100 °C, positive pressure 600 to 900 mbar, vacuum pressure -800 to -1000 mbar, evaporation 10 to 20 seconds. The preparation method according to item 11. 16. Use of the liquid composition of Compound I prepared by the method according to any one of items 1 to 15 in a myocardial perfusion PET contrast agent.

[0008] Also, the present application provides a liquid composition containing Compound I, and the specific technical solution is as follows. 1. A liquid composition of Compound I comprising one or more selected from vitamin C, sodium vitamin C, and gentisic acid, wherein Compound I is 2-tert-butyl-4-chloro-5-((3-((4-((2-(2-fluoro 18 F]ethoxy)ethoxy)methyl)-1H-1,2,3-triazol)-1-yl)methyl)benzyloxy)pyridazin-3(2H)-one. 2. The liquid composition according to item 1, wherein the ratio of the vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.02 to 0.2. 3. The liquid composition according to item 2, wherein the ratio of the vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15. 4. The liquid composition according to item 1, wherein the ratio of the gentisic acid concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.02 to 0.2. 5. The liquid composition according to item 4, wherein the ratio of the gentisic acid concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15. 6. The liquid composition according to item 1, wherein the ratio of the sodium vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 1. 7. The liquid composition according to item 6, wherein the ratio of the sodium vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.1 to 1. 8. The liquid composition according to item 1, wherein the sodium vitamin C is 2 to 10 parts by weight with respect to 1 part by weight of the vitamin C. 9. The liquid composition according to item 8, wherein the sodium vitamin C is 3 to 10 parts by weight with respect to 1 part by weight of the vitamin C. 10. The liquid composition according to item 1, wherein the gentisic acid is 0.1 to 5 parts by weight with respect to 1 part by weight of the vitamin C. 11. The liquid composition according to item 1, wherein the ratio of the sodium vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.1 to 1, and the ratio of the gentisic acid concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15. 12. The liquid composition according to item 1, wherein the ratio of the vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15, and the ratio of the sodium vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.1 to 1. 13. The liquid composition according to item 1, wherein the ratio of the vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15, and the ratio of the gentisic acid concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15. 14. The liquid composition according to item 1, wherein the ratio of the vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15, the ratio of the sodium vitamin C concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.1 to 1, and the ratio of the gentisic acid concentration (mg / mL) to the activity concentration of Compound I (mCi / mL) is 0.04 to 0.15. 15. The liquid composition according to item 1, further comprising a pharmaceutically acceptable carrier. 16. A method for preparing the liquid composition according to item 1, comprising dissolving one or more selected from vitamin C, sodium vitamin C, and gentisic acid in a solvent containing Compound I. 11. The genisteic acid is 0.1 to 3 parts by weight with respect to 1 part by weight of the vitamin C1. The liquid composition according to item 10. 12. The liquid composition according to item 1, wherein the formulation of the liquid composition further contains an aqueous solution of polyethylene glycol and / or ethanol. 13. The concentration of the polyethylene glycol is 0.1 to 0.3 g / mL, preferably 0.1 to 0.2 g / mL. The liquid composition according to item 12. 14. The polyethylene glycol is polyethylene glycol 400. The liquid composition according to item 12. 15. The ethanol is 5 to 20 mL with respect to 100 mL of water. The liquid composition according to item 12. 16. Dissolve one or more of vitamin C, sodium ascorbate, and genisteic acid in water, stir, recover Compound I, and mix them to obtain a liquid composition of Compound I. The method for preparing the liquid composition according to any one of items 1 to 15. 17. Dissolve one or more of vitamin C, sodium ascorbate, and genisteic acid, a polyethylene glycol-based substance and / or ethanol in water, stir, recover Compound I, and mix them to obtain the liquid composition of the said Compound I. The method for the liquid composition according to item 16. 18. Use of the liquid composition according to any one of items 1 to 15, and the liquid composition of Compound I prepared by the method according to item 16 or 17 in the preparation of a myocardial perfusion PET contrast agent.

[0009] Furthermore, the present application provides a reagent kit for the automatic preparation of a liquid composition of Compound I, and the reagent kit includes a reaction bottle and a formulation bottle. The reaction bottle includes reaction bottle R1, reaction bottle R2, and reaction bottle R3. The reaction bottle R1 is 18 Used for containing an eluent for eluting F ions. The reaction bottle R2 is 18 used to contain a solution of Compound I precursor for the nucleophilic substitution reaction of F ions, The reaction bottle R3 is used to contain a reagent used to dilute the crude product and rinse the reaction system, The formulation bottle includes formulation bottle P1 and formulation bottle P2, The formulation bottle P1 is 18 used to transfer the reaction product after the nucleophilic substitution reaction of F ions, The formulation bottle P2 is 18 used to contain a substance used to stabilize the reaction product after the nucleophilic substitution reaction of F ions.

[0010] Furthermore, the reaction bottle R1 contains aminopolyether, potassium carbonate, water for injection, and acetonitrile. Preferably, the concentration of the aminopolyether is 5 - 40 mg / mL, the concentration of the potassium carbonate is 1.5 - 20 mg / mL, and the volume ratio of acetonitrile to water is (0.25 - 19):1. Alternatively, the reaction bottle R2 18 contains an acetonitrile solution of Compound I precursor for the nucleophilic substitution reaction of F ions. Preferably, the concentration of the Compound I precursor in the acetonitrile solution is 1 - 10 mg / mL. Alternatively, the reaction bottle R3 contains absolute ethanol.

[0011] Furthermore, the formulation bottle P1 contains polyethylene glycol. Preferably, the concentration of the polyethylene glycol is 0.05 - 0.3 g / mL. Alternatively, the formulation bottle P2 18 contains one or more of vitamin C, sodium ascorbate, and gentisic acid used to stabilize the reaction product after the nucleophilic substitution reaction of F ions. Preferably, the vitamin C concentration is 2 - 16 mg / mL, the sodium ascorbate is 15 - 40 mg / mL, and the gentisic acid concentration is 2 - 16 mg / mL.

[0012] Compared with the existing technology, the beneficial effects of this application are as follows. (1) This application optimizes the experimental process plan, changes the dosage of Compound I precursor, shortens the reaction time to achieve the same labeling rate, increases the initial 18 radioactivity of F ions, and increases the yield by increasing the labeling rate. The process parameters and process flow are clear and specific, applicable to large-scale batch production, and can meet the needs of automation. (2) In the optimized technical solution of this application, a radiolysis inhibitor is used in the mobile phase of the purification process to avoid product loss due to radiolysis during the purification and formulation processes, thereby improving the yield. Furthermore, in the formulation process of this application, a radiolysis inhibitor is used, so the stability of the product can be ensured. (3) This application optimizes the purification process, removes the C18 cartridge, and uses ethanol / water system instead of acetonitrile / water system as the mobile phase. This shortens the time and improves the radiochemical purity and stability of the product. (4) The reagent kit of this application provides convenience, cost savings, and accurate control for the automatic preparation of the liquid composition of Compound I. (5) In the prior art, it is common to prepare samples temporarily, and each time the preparation is completed, the inspector needs to conduct inspections before shipment. This results in a large workload, low production efficiency, and high costs in the production process. However, after this application is prepared into a reagent kit, it can be directly used in an automated device to directly prepare the liquid composition of Compound I. The process is simple, the operation is easy, and the quality of the final product can be controlled. Also, under conditions that meet GMP, management can be standardized according to the established manufacturing process requirements, and reagent kits that meet the expected quality standards can be continuously and stably produced. In this way, the overall process flow is simplified and the production cost is significantly reduced. Details of the invention

[0013] The chemical name of Compound I is 2-tert-butyl-4-chloro-5-((3-((4-((2-(2-fluoro 18 F]ethoxy)ethoxy)methyl))-1H-1,2,3-triazol-1-yl)methyl)benzyloxy)pyridazin-3(2H)-one. The chemical structural formula is as follows.

[0014]

Chemical Structure

[0015] Molecular formula: C 23 H 29 Cl 18 FN5O4 Molecular weight: 492.97

[0016] The mechanism of action of Compound I as a myocardial perfusion PET imaging agent is as follows. When Compound I enters cardiomyocytes, it immediately interacts with respiratory chain complex I (MC-I) in mitochondria and accumulates in the myocardium for a long time. Preliminary animal research data show that the uptake in the heart is high and the uptake in the liver is low 15 minutes after injection, and a good heart-to-liver ratio is maintained 60 minutes after injection, indicating the potential for good myocardial perfusion imaging.

[0017] In this application, the liquid composition of Compound I or Compound I is used as a myocardial perfusion PET imaging agent.

[0018] Compound I precursor: The chemical name is methyl 2-(2-((1-(3-(((1-(tert-butyl))-5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)oxy))methyl)benzyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethyl-4-methylbenzenesulfonate, and the chemical structural formula is as follows.

[0019]

Chemical Structure

[0020] Molecular formula: C30 H 36 ClN5O7S Molecular weight: 646.16

[0021] Aminopolyether (K 222 ) is a three - bridged crown ether molecule with a cryptoid cavity, a typical azacryptand, and a type of cryptand. Due to its unique coordination properties, the azacryptand can appropriately select cations that complex transition metals and heavy metals, and the resulting complexes are more stable and have lipophilicity and hydrophilicity, thus having excellent research prospects.

[0022] A typical synthesis method of aminopolyether (K 222 ) in the prior art is the highly diluted method proposed by Lehn et al., which is one of the typical non - template ion synthesis methods. The specific procedure is as follows. The starting materials 1,8 - diamino - 3,6 - dioxaoctane and 1,8 - diacyl chloride - 3,6 - dioxaoctane are dissolved in a large amount of benzene solvent, heated and reacted for 8 hours, then reduced with lithium aluminum hydride for 24 hours, separated by column chromatography, and recrystallized to obtain aminopolyether (K 222 ). This method requires a large amount of solvents such as benzene, has a long synthetic route, complex operations, low yield, and low economic benefits. In addition to the highly diluted method, another typical synthesis method of aminopolyether (K 222 ) is the synthesis method proposed by Kulstad and Malmsten. In this synthesis method, using Na2CO3 etc. as a template, an aminopolyether (K 222 ) sodium iodide complex is obtained in acetonitrile, and then this complex is decomplexed with resin to obtain aminopolyether (K 222To obtain it. As the specific procedure, 1,2-bis(2-iodoethoxy)ethane as the raw material and benzylamine are refluxed in an acetonitrile solution for 3 days, and an intermediate is obtained through post-treatment. This intermediate is recrystallized with acetone, and after filtration, an NaI complex is obtained. This complex is decomplexed with a cation exchange resin and an anion exchange resin respectively under acidic conditions to prepare an aminopolyether (K 222 ) is prepared. This method has a simple apparatus, low solvent consumption, and relatively mild reaction conditions. However, as a result of research, the applicant has discovered that in the method of decomplexing using an ion exchange resin, when the sodium ion content is reduced to a certain amount, the decomplexing cannot proceed and the yield decreases.

[0023] In this application, the applicant has discovered that in the existing preparation method of Compound I, a large batch of liquid composition of Compound I that meets the quality requirements cannot be obtained, and a product with a high activity concentration cannot be obtained either.

[0024] In this application, a large batch refers to a product with a high total activity, and generally can refer to a product with a total activity exceeding 1 Ci, that is, 37 GBq. A product with a high activity concentration generally can refer to a product with an activity concentration exceeding 50 mCi / mL, that is, 1850 MBq / mL.

[0025] In this application, the labeling rate refers to 18 conducting a labeling reaction between [¹⁸F] and the reaction precursor, 18 [¹⁸F] replacing the leaving group in the precursor and converting it into the final labeled product. Since the labeled product contains 18 [¹⁸F], the labeling rate is defined as the ratio of the activity of the labeled product to the total 18 [¹⁸F] activity involved in the reaction.

[0026] In this application, the yield refers to 18 the ratio of the activity of the liquid composition of Compound I, which is the final product, to the initial

[0027] For example, in the prior art, high-performance liquid chromatography (HPLC) is used for purification, and the mobile phase used in this purification step is a mixed system of acetonitrile and water. The applicant has found that 18 when the activity of F is increased, the crude product of compound I obtained increases. If the existing method is used as it is, the difficulty of purification increases, and 18 it has been found that when the initial F activity is high, the labeling rate decreases. At the same time, the yield calculated after purification also decreases, making it impossible to obtain a large batch of liquid composition of compound I that meets the quality requirements. From this, it can be seen that using a mixed system of acetonitrile and water as the mobile phase to purify the crude product of compound I results in very low labeling rates and yields. 18 Furthermore, it can be seen that simply increasing the initial F ion activity using the reaction and purification conditions of the existing technology cannot produce the required high-quality and large-batch products.

[0028] In addition, the applicant has also found that when using a mixed system of acetonitrile and water as the mobile phase to purify the crude product of compound I, it is necessary to use a C18 cartridge to remove organic solvents such as acetonitrile in the HPLC mobile phase. However, since compound I is highly sensitive to radiolysis, it decomposes during purification. During the purification with a C18 cartridge, radioactive substances are concentrated into a very small volume, causing radiolysis. After radiolysis, the yield decreases. Furthermore, the radiochemical purity and stability of the final compound I product cannot meet the requirements, and the preparation time becomes longer. A C18 cartridge is an octadecyl-bonded silica gel cartridge that can be used for the concentration and removal of organic solvents such as acetonitrile.

[0029] In order to obtain a large batch of liquid composition of compound I that meets the quality requirements, increase the labeling rate, improve the radiochemical purity and stability of the final product, and shorten the time of the entire reaction process, the applicant has created a technical solution that can solve the problems of low labeling rate, low radiochemical purity, and low stability through repeated verification through multiple studies.

[0030] This application provides a method for preparing Compound I, and the synthetic route is as follows.

[0031]

Chemical formula

[0032] Activated 18 Mix the F⁻ ions with an acetonitrile solution containing the precursor of Compound I, and carry out a nucleophilic substitution reaction under the action of a catalyst to produce a crude product containing Compound I.

[0033] The method for preparing the above Compound I provided by this application purifies the crude product containing Compound I using high-performance liquid chromatography (HPLC). The mobile phase used in the purification step by high-performance liquid chromatography contains ethanol and water instead of the original acetonitrile and water system. When using the acetonitrile and water system, a C18 cartridge is required to remove the organic solvent, resulting in high activity and radiolysis, which cannot meet the production needs.

[0034] In the method for preparing the above Compound I provided by this application, sodium ascorbate (VcNa) and / or vitamin C (Vc) and / or genisteic acid, which are radiolysis inhibitors, are added to the mobile phase used in the purification step by high-performance liquid chromatography. Sodium ascorbate, vitamin C, and genisteic acid have a radiolysis prevention effect. During the HPLC purification process, radioactive 18 To avoid radiolysis due to the enrichment of F⁻ ions, adding Vc and / or VcNa and / or genisteic acid to the mobile phase can reduce the radiolysis of Compound I during the column purification process.

[0035] The above method for preparing Compound I provided by this application improves the radiochemical purity of the liquid composition of Compound I, removes the purification step using a C18 cartridge, and reduces the radiolysis on the C18 cartridge.

[0036] In the preparation method of the present application, after removing the C18 cartridge, the step of removing the solvent is omitted, and ethanol for injection can be used as the mobile phase. Further, when ethanol is used as the mobile phase, the risk of high-risk solvents such as acetonitrile remaining in the liquid composition is reduced.

[0037] In some specific embodiments of the present application, in the purification step by high-performance liquid chromatography, in the mobile phase, the ethanol is 0.2 to 2 parts by volume with respect to 1 part by volume of water, and preferably, in the mobile phase, the ethanol is 0.4 to 1 part by volume with respect to 1 part by volume of water.

[0038] For example, with respect to 1 part by volume of water, the ethanol can be 0.2 part by volume, 0.3 part by volume, 0.4 part by volume, 0.5 part by volume, 0.6 part by volume, 0.7 part by volume, 0.8 part by volume, 0.9 part by volume, 1 part by volume, 1.1 parts by volume, 1.2 parts by volume, 1.3 parts by volume, 1.4 parts by volume, 1.5 parts by volume, 1.6 parts by volume, 1.7 parts by volume, 1.8 parts by volume, 1.9 parts by volume, 2 parts by volume, or any range therebetween.

[0039] In some specific embodiments of the present application, in the purification step by high-performance liquid chromatography, in the mobile phase, the added amount of sodium ascorbate is 0 to 20 mg / mL, and preferably, the added amount of sodium ascorbate is 0.2 to 10 mg / mL. Here, the added amount of sodium ascorbate can be 0, 2, 5, 8, 10, 12, 15, 20 mg / mL, or any range therebetween.

[0040] In some specific embodiments of the present application, in the purification step by high-performance liquid chromatography, in the mobile phase, the added amount of ascorbic acid is 0 to 10 mg / mL, and preferably, the added amount of ascorbic acid is 0.1 to 5 mg / mL. The added amount of ascorbic acid can be 0, 0.1, 0.5, 1, 2, 5, 8, 9, 10 mg / mL, or any range therebetween.

[0041] In some specific embodiments of the present application, in the purification step by high performance liquid chromatography, in the mobile phase, the addition amount of the gentiopic acid is 0 to 10 mg / mL, preferably, the addition amount of the gentiopic acid is 0.1 to 5 mg / mL, The addition amount of the gentiopic acid can be 0, 0.1, 0.5, 1, 2, 5, 8, 9, 10 mg / mL, or any range therebetween.

[0042] In some embodiments of the present application, the chromatography column is a silica gel column, preferably a reverse phase C18 silica gel chromatography column, more preferably an XBridge BEH C18 OBD Prep column. This chromatography column is selected because it has acid and alkali resistance and good resolution.

[0043] In some embodiments of the present application, isocratic elution is used in the purification step, and the elution flow rate of the mobile phase is 3 to 6 mL / min. For example, the elution flow rate of the mobile phase can be 3, 4, 5, 6 mL / min, or any range therebetween.

[0044] In some embodiments of the present application, before the purification step by high performance liquid chromatography, a step of performing a nucleophilic substitution reaction is also included. In the nucleophilic substitution reaction, activated 18 F ions are mixed with the precursor-containing solution of Compound I to perform a nucleophilic substitution reaction to produce a crude product containing Compound I.

[0045] Activated 18 The nucleophilic substitution reaction between the F ions and the precursor of Compound I can be carried out by a method known to those skilled in the art.

[0046] In some embodiments of the present application, in the nucleophilic substitution reaction, the reaction solvent is an aprotic polar solvent, and the dosage of the Compound I precursor / 18The range of the ratio of F ion activity is (0.5 to 8):1, the reaction temperature is 90 to 140 °C, the reaction time is 5 to 60 minutes, it is a sealed reaction, and the unit of the dosage of Compound I precursor is mg. 18 The unit of F ion activity is Ci.

[0047] Here, the dosage of Compound I precursor / 18 The range of the ratio of F ion activity can be 0.5:1, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or any range therebetween, and the unit of the dosage of Compound I precursor is mg. 18 The unit of F ion activity is Ci. The reaction temperature can be 90, 100, 110, 120, 130, 140 °C, or any range therebetween. The reaction time can be 5, 15, 20, 35, 45, 60 minutes, or any range therebetween.

[0048] In some embodiments of the present application, the dosage of the reaction solvent is 0.2 to 5 mL, and the dosage of the Compound I precursor is 0.8 to 20 mg, preferably 7 to 20 mg. Here, the dosage of the Compound I precursor can be 0.8, 2, 6, 7, 8, 12, 18, 20 mg, or any range therebetween. In some embodiments of the present application, the aprotic polar solvent is one or more of acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, and 2-methyl-2-butanol.

[0049] In some embodiments of the present application, before the nucleophilic substitution reaction step, 18 an F ion preparation step is also included. 18 The F ion preparation step 18 includes the preparation of an F ion solution, 18 the concentration and elution of F ions, and 18 further includes the activation of F ions. Here, 18 in the preparation of the F ion solution, an F ion solution is prepared by an accelerator. 18 The F ion solution is prepared, 18 In the concentration of F ions, the F ion solution prepared above is concentrated through an anion exchange cartridge, 18 and in the elution of F ions, a cryptand and an alkali metal salt catalyst solution are used to elute the F ions, 18 and in the activation of F ions, the temperature is programmed to dry the solvent with nitrogen or other inert gases, 18 activate the F ions, and obtain the activated F ions. 18 In some embodiments of the present application, in the step of preparing the F ion solution, 18 water containing O is transferred to the target position of the accelerator, the accelerator is started to generate a proton beam, 18 and the water containing O is collided to generate a solution containing F ions.

[0050] In some embodiments of the present application, 18 in the step of concentrating the F ions, the anion exchange cartridge is a Sep-Pak Accell Plus QMA Carbonate Plus Light Cartridge, specifically a tetraalkylammonium salt anion exchange cartridge. 18 18 18 18 18 18

[0051] In the present application, 18 the initial activity of F, also referred to as the F ion activity, is the activity of the F ions measured using an activity meter, which is obtained by starting the accelerator to generate a proton beam and colliding the water containing oxygen

[0052]

[0053] 18 O] to generate a solution containing F ions. 18 In the present application, the initial activity of F refers to starting the accelerator to generate a proton beam and colliding the water containing oxygen 18 O] to generate a solution containing F ions, 18 and the activity of the F ions measured using an activity meter.

[0053] In the present application, 18 the initial activity of F refers to starting the accelerator to generate a proton beam and colliding the water containing oxygen 18 O] to generate a solution containing F ions, 18It refers to the concentration that can be immediately detected after generating a solution containing F ions. "Immediate detection" refers to a reasonable detection time that can be controlled by those skilled in the art, such as within 10 minutes after generation. Also, when the standing time after generation changes, 18 the initial activity of F changes to a certain extent, but usually, those skilled in the art will understand that the error range is within ±10%.

[0054] In some embodiments of the present application, the 18 F ion activity is 0.045 Ci to 11 Ci, preferably 3.15 Ci to 11 Ci. For example, 18 the F ion activity can be 0.045 Ci, 0.05 Ci, 1 Ci, 3 Ci, 3.15 Ci, 5 Ci, 6 Ci, 8 Ci, 9 Ci, 10 Ci, 11 Ci, or any range therebetween.

[0055] In some embodiments of the present application, the 18 In the elution step of F ions, in the catalyst solution of cryptand and alkali metal salt, the dosage of cryptand is 5 to 40 mg, and the dosage of alkali metal salt is 1.5 to 20 mg. The dosage of cryptand in the solution can be 5, 8, 10, 15, 20, 40 mg, or any range therebetween, and the dosage of alkali metal salt in the solution can be 1.5, 3, 5, 10, 20 mg, or any range therebetween.

[0056] In some embodiments of the present application, the cryptand is 4,7,13,16,21,24 - hexaoxa - 1,10 - diazabicyclo[8,8,8]hexacosane (cryptofix - 2.2.2, aminopolyether), and the alkali metal salt is one or more of K2CO3, Na2CO3, Cs2CO3, KHCO3, and NaHCO3.

[0057] In some embodiments of the present application, the catalyst solution is selected from a mixed solvent system of acetonitrile and water, and the volume ratio of acetonitrile to water is (0.2 to 10):1. For example, the volume ratio of acetonitrile to water can be 0.2:1, 1:1, 2:1, 4:1, 7:1, 10:1, or within the range therebetween. The volume of the mixed solvent of acetonitrile and water is 0.3 to 2 mL.

[0058] In some embodiments of the present application, 18 In the activation step of F ions, the activation temperature is 80 to 130 °C. Here, the programmed temperature of the temperature includes the following steps: 100 to 120 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation 60 to 120 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation 150 to 200 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -60 to -100 mbar, evaporation 10 to 30 seconds; 100 to 120 °C, positive pressure 800 to 1200 mbar, vacuum pressure -800 to -1000 mbar, evaporation 80 to 120 seconds; 80 to 100 °C, positive pressure 400 to 600 mbar, vacuum pressure -800 to -1000 mbar, evaporation 100 to 120 seconds; 80 to 100 °C, positive pressure 600 to 900 mbar, vacuum pressure -800 to -1000 mbar, evaporation 10 to 20 seconds.

[0059] The preparation method of the above compound I provided by the present application can change the dosage of the compound I precursor, shorten the reaction time to achieve the same labeling rate, increase the labeling rate, thereby increasing the yield, and obtain a compound I product. The process parameters and process flow are clear and specific, can be applied to large-scale batch production, and can meet the needs of automation. Furthermore, a radiation decomposition inhibitor is used in the mobile phase of the purification process to avoid product loss due to radiation decomposition during the purification and formulation processes, thereby improving the yield. Furthermore, in the formulation process of the present application, a radiation decomposition inhibitor is used, so the stability of the product can be ensured.

[0060] The increase in the reaction solvent is based on the fact that Compound I required after the completion of the reaction is a lipophilic substance. After dilution with water, when the proportion of the organic phase increases, the residue of Compound I in the reaction bottle decreases, increasing the yield.

[0061] In some embodiments of the present application, the preparation of Compound I includes the following steps: 1) 18 Preparation of F-ion solution Oxygen 18 2-3 g of water containing 18 O] is transferred to the target position of the accelerator, and the accelerator is started to generate a proton beam to collide with the water containing 18 O], 2) 18 Concentration of F-ion The 18 F-ion solution prepared above is passed through an anion exchange solid phase extraction cartridge (QMA cartridge of Waters brand, and the QMA cartridge is preferably activated with 1 mol / L NaHCO3), 18 and the F-ion is concentrated on the QMA cartridge. 3) 18 Elution of F-ion Using a cryptand and an alkali metal salt catalyst solution, 18 the F-ion is eluted into the reaction bottle. Specifically, 222 5-40 mg of 18 F / 222 complex is eluted into the reaction bottle. 4) 18 Activation of F-ion The 18 F-ion eluted in step 3) is heated to 80-130 °C under a nitrogen or inert gas flow to dry the solvent, and 18 activated 5) 18 Nucleophilic substitution reaction of F-ion Add 0.2 - 5 mL of an acetonitrile solution of the compound I precursor to a reaction bottle. The dosage of the compound I precursor is 7 - 20 mg. Heat it to 90 - 140 °C under sealed conditions and react for 5 - 60 minutes. The nucleophilic substitution reaction between the compound I precursor and K 18 F / K 222 proceeds to obtain a crude product containing compound I. 6) Purification by high - performance liquid chromatography Load the crude product containing compound I into a sample loop. After extracting a certain amount of water and rinsing the reaction bottle, load it into the sample loop and purify it according to the following chromatography conditions. Column: Reverse - phase C18 silica gel column Mobile phase: A mixed system of ethanol and water. Here, the volume ratio of ethanol to water is (0.2 - 2):1, and the mobile phase contains 0 - 20 mg / mL of sodium ascorbate and / or 0 - 10 mg / mL of ascorbic acid and / or 0 - 10 mg / mL of gentisic acid; Flow rate: 3 - 6 mL / min Detector: Radioactivity detector Monitor and track the radioactive signal, and collect the radioactive main peak of compound I into a transfer bottle. After purification, a purified product of compound I is obtained.

[0062] In some embodiments of the present application, the mobile phase may contain 0 - 20 mg / mL of sodium ascorbate and / or 0 - 10 mg / mL of ascorbic acid.

[0063] In some embodiments of the present application, the mobile phase may contain 0 - 20 mg / mL of sodium ascorbate and / or 0 - 10 mg / mL of gentisic acid.

[0064] In some embodiments of the present application, the mobile phase may contain both ascorbic acid and gentisic acid, or may contain only one of ascorbic acid and gentisic acid.

[0065] This application also provides a liquid composition of Compound I above. The compound I is collected in a transfer bottle with a pre-added formulation, and the liquid composition of compound I is mixed with the HPLC main peak mobile phase to obtain the liquid composition of compound I. The liquid composition of compound I contains one or more of vitamin C, sodium ascorbate, and gentisic acid.

[0066] In some embodiments of the present application, the ratio of vitamin C concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.02 - 0.2. Preferably, the ratio of vitamin C concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.04 - 0.15. For example, the ratio of vitamin C concentration (mg / mL) / compound I activity concentration (mCi / mL) can be 0.02, 0.04, 0.06, 0.08, 0.1, 0.12, 0.14, 0.16, 0.18, 0.2, or any range therebetween.

[0067] In some embodiments of the present application, the ratio of gentisic acid concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.02 - 0.2. Preferably, the ratio of gentisic acid concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.04 - 0.15. For example, the ratio of gentisic acid concentration (mg / mL) / compound I activity concentration (mCi / mL) can be 0.02, 0.04, 0.06, 0.08, 0.1, 0.12, 0.14, 0.16, 0.18, 0.2, or any range therebetween.

[0068] In some embodiments of the present application, the ratio of sodium ascorbate concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.04 - 1. Preferably, the ratio of sodium ascorbate concentration (mg / mL) / compound I activity concentration (mCi / mL) is 0.1 - 1. For example, the ratio of sodium ascorbate concentration (mg / mL) / compound I activity concentration (mCi / mL) can be 0.04, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, or any range therebetween.

[0069] In some embodiments of the present application, the sodium ascorbate is 2 to 10 parts by weight based on 1 part by weight of vitamin C. Preferably, based on 1 part by weight of vitamin, the sodium ascorbate is 3 to 10 parts by weight. For example, the sodium ascorbate may be 2 parts by weight, 3 parts by weight, 4 parts by weight, 5 parts by weight, 6 parts by weight, 7 parts by weight, 8 parts by weight, 9 parts by weight, 10 parts by weight, or any range therebetween based on 1 part by weight of vitamin C.

[0070] In some embodiments of the present application, the gentiopic acid is 0.1 to 5 parts by weight based on 1 part by weight of vitamin C, and preferably, the gentiopic acid is 0.1 to 3 parts by weight based on 1 part by weight of vitamin C. For example, based on 1 part by weight of vitamin C, the gentiopic acid may be 0.1 part by weight, 0.5 part by weight, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, 3 parts by weight, 3.5 parts by weight, 4 parts by weight, 4.5 parts by weight, 5 parts by weight, or any range therebetween.

[0071] In some embodiments of the present application, the liquid composition of the compound I may contain both vitamin C and gentiopic acid, or may contain only one of vitamin C and gentiopic acid.

[0072] In some embodiments of the present application, the liquid composition of the compound I also contains an aqueous solution of polyethylene glycol and / or ethanol, the polyethylene glycol concentration is 0.1 to 0.3 g / mL, and preferably, the polyethylene glycol concentration is 0.1 to 0.2 g / mL. For example, the polyethylene glycol concentration is 0.1, 0.2, 0.3 g / mL, or any range therebetween.

[0073] In some embodiments of the present application, the ethanol concentration in the aqueous ethanol solution is 0.05 to 0.20 mL / mL, and preferably, the ethanol concentration in the aqueous ethanol solution is 0.05 to 0.15 mL / mL. The ethanol concentration in the aqueous ethanol solution is 0.05, 0.10, 0.15, 0.20 mL / mL, or any range therebetween.

[0074] In some embodiments of the present application, the polyethylene glycol is polyethylene glycol 400.

[0075] The present application also provides a method for preparing a liquid composition of Compound I, in which one or more of vitamin C, sodium vitamin C, and gentiopic acid, a polyethylene glycol-based substance and / or ethanol are dissolved in water and stirred to obtain a liquid composition formulation of Compound I.

[0076] In some embodiments of the present application, purified Compound I is collected in a prescription bottle, in which the above liquid composition is pre-added, and the process is completed when the liquid composition is mixed with the main peak mobile phase in purification by high performance liquid chromatography.

[0077] The present application also provides the use of the liquid composition of the above Compound I in a myocardial perfusion PET contrast agent.

[0078] The present application provides a reagent kit for the automatic preparation of a liquid composition of Compound I, the reagent kit includes a reaction bottle and a prescription bottle, the reaction bottle includes reaction bottles R1, R2, and R3, and reaction bottle R1 is 18 used to contain an eluent for F ion elution, reaction bottle R2 is 18 used to contain a solution of a Compound I precursor for the nucleophilic substitution reaction of F ions, reaction bottle R3 is used to contain reagents for diluting the crude product and rinsing the reaction system, the prescription bottle includes prescription bottles P1 and P2, and prescription bottle P1 is 18 used to transfer the reaction product after the nucleophilic substitution reaction of F ions, and prescription bottle P2 is 18 used to contain a substance for stabilizing the reaction product after the nucleophilic substitution reaction of F ions.

[0079] In some embodiments of the present application, the reaction bottle R1 contains aminopolyether, potassium carbonate, water for injection, and acetonitrile. Preferably, the concentration of the aminopolyether is 5 to 40 mg / mL, the concentration of the potassium carbonate is 1.5 to 20 mg / mL, and the volume ratio of acetonitrile to water is (0.25 to 19):1. For example, the concentration of the aminopolyether can be 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, 21 mg / mL, 22 mg / mL, 23 mg / mL, 24 mg / mL, 25 mg / mL, 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 31 mg / mL, 32 mg / mL, 33 mg / mL, 34 mg / mL, 35 mg / mL, 36 mg / mL, 37 mg / mL, 38 mg / mL, 39 mg / mL, 40 mg / mL, or any range therebetween. The concentration of the potassium carbonate can be 1.5 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, or any range therebetween. The volume ratio of acetonitrile to water can be 0.25:1, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, or any range therebetween.

[0080] In some embodiments of the present application, the reaction bottle R2 is 18Contains an acetonitrile solution of Compound I precursor for F-ion nucleophilic substitution reaction. Preferably, the concentration of Compound I precursor in the acetonitrile solution is 1-10 mg / mL. For example, the concentration of Compound I precursor in the acetonitrile solution can be 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, or any range therebetween.

[0081] In some embodiments of the present application, the reaction bottle R3 contains absolute ethanol.

[0082] In some embodiments of the present application, the formulation bottle P1 contains polyethylene glycol. Preferably, the concentration of the polyethylene glycol is 0.05-0.3 g / mL. For example, the concentration of the polyethylene glycol can be 0.05 g / mL, 0.1 g / mL, 0.15 g / mL, 0.2 g / mL, 0.25 g / mL, 0.3 g / mL, or any range therebetween.

[0083] In some embodiments of the present application, the formulation bottle P2 18Contains one or more of vitamin C, sodium ascorbate, and gentisic acid used to stabilize the reaction product after the nucleophilic substitution reaction of F ions, where vitamin C, sodium ascorbate, and gentisic acid are dissolved in water before use. Preferably, the concentration of the vitamin C is 2 to 16 mg / mL, the sodium ascorbate is 15 to 40 mg / mL, and the concentration of the gentisic acid is 2 to 16 mg / mL. For example, the concentration of the vitamin C can be 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, or any range therebetween. The sodium ascorbate can be 15 mg / mL, 16 mg / mL, 17 mg / mL, 18 mg / mL, 19 mg / mL, 20 mg / mL, 21 mg / mL, 22 mg / mL, 23 mg / mL, 24 mg / mL, 25 mg / mL, 26 mg / mL, 27 mg / mL, 28 mg / mL, 29 mg / mL, 30 mg / mL, 31 mg / mL, 32 mg / mL, 33 mg / mL, 34 mg / mL, 35 mg / mL, 36 mg / mL, 37 mg / mL, 38 mg / mL, 39 mg / mL, 40 mg / mL, or any range therebetween. The concentration of the gentisic acid can be 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, 12 mg / mL, 13 mg / mL, 14 mg / mL, 15 mg / mL, 16 mg / mL, or any range therebetween.

[0084] The reaction bottles R1, R2, R3, the formulation bottles P1, and the formulation bottle P2 can be configured according to the actual needs as long as their concentration ranges meet the needs.

[0085] This application provides a general and / or specific description of the materials used in the tests and the test methods. In the following examples, unless otherwise specifically indicated, % represents weight %, i.e., weight percent. When the manufacturer of a reagent or instrument used is not indicated, they are all commercially available conventional reagent products. Table 1 shows the sources of the raw materials used in the examples.

[0086]

Table 1

Examples

[0087] An automated device is a device with the model number AllinOne from Trasis. The power unit of this device is high-purity nitrogen and an electric syringe rotor, which can provide a vacuum system and is equipped with an HPLC purification system. Since this process uses an automated device installed in a radiation shielding box, it can protect the operator from radiation hazards, increase the working dose, and with computer control, the control of the process steps becomes more accurate, the reproducibility is improved, and human errors are reduced.

[0088] Example 1 Preparation of Compound I 1) 18 Preparation of F-ion solution Oxygen 18 2 g of water containing 18 O] was transferred to the target position of the accelerator, and the accelerator was started to generate a proton beam to collide with the water containing 18 O], 18 and a solution containing F ions was generated. In this example, a large amount of starting material 18 F was used, and the initial activity of 2) 18 Concentration of F ions The 18 F-ion solution prepared above was passed through an anion exchange solid phase extraction cartridge (a QMA cartridge of the Waters brand, preferably a QMA cartridge activated with 1 mol / L NaHCO3), and then 18F ions were concentrated on the QMA cartridge. 3) 18 Elution of F ions Using a cryptand and an alkali metal salt catalyst solution 18 F ions were eluted into the reaction bottle. Specifically, K 222 15 mg (dissolved in 0.9 mL of acetonitrile) was mixed with 1.5 mg of K2CO3 (dissolved in 0.1 mL of water) to prepare a mixed solvent system of acetonitrile and water (volume ratio of acetonitrile to water is 9:1) solution (that is, reaction bottle R1, for its formulation components and preparation, refer to the preparation of reaction bottle R1), the above QMA cartridge was eluted, and K 18 F / K 222 The complex was eluted into the reaction bottle. 4) 18 Activation of F ions The F ions eluted in step 3) 18 were programmed to heat to 80 - 130 °C under a nitrogen gas flow, and the solvent was dried by blowing air to obtain activated 18 F ions. 5) 18 Nucleophilic substitution reaction of F ions 3 mL of an acetonitrile solution containing compound I precursor (that is, reaction bottle R2, for its formulation components and preparation, refer to the preparation of reaction bottle R2) was added to the reaction bottle. The dosage of compound I precursor was 12 mg, heated to 100 °C under sealed conditions, and reacted for 10 minutes. Compound I precursor and K 18 F / K 222 underwent a nucleophilic substitution reaction to obtain a crude product containing compound I. 6) Purification by high performance liquid chromatography The crude product containing compound I was loaded into the sample loop, 2.5 mL of water was extracted to rinse the reaction bottle, then loaded into the sample loop, and purified according to the following chromatography conditions. Column: XBridge BEH C18 OBD Prep column, 130A, 5 μm, 10 × 250 mm Mobile phase: A mixed system of ethanol and water. Here, the volume ratio of ethanol to water is 0.6:1, and the mobile phase contains 5 mg / mL of sodium ascorbate and 1 mg / mL of vitamin C (the mobile phase can be obtained through prescription bottle P2. For its formulation components and preparation, refer to the preparation of prescription bottle P2); Flow rate: 4 mL / min Detector: Radioactivity detector The radioactive signal was monitored and traced, and the radioactive main peak of Compound I was collected into the transfer bottle. After purification, a purified product of Compound I was obtained.

[0089] Example 2 The difference between Example 2 and Example 1 is 5) 18 In the nucleophilic substitution reaction of F ions, the dosage of the Compound I precursor is only 20 mg.

[0090] Example 3 The difference between Example 3 and Example 2 is 1) 18 In the preparation of the F ion solution 18 The initial activity of F is only 6 Ci.

[0091] Example 4 The difference between Example 4 and Example 2 is 1) 18 In the preparation of the F ion solution 18 The initial activity of F is only 10 Ci.

[0092] Example 5 The difference between Example 5 and Example 4 is 6) In the purification by high performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 2:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of vitamin C.

[0093] Example 6 The difference between Example 6 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 1:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of ascorbic acid.

[0094] Example 7 The difference between Example 7 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate.

[0095] Example 8 The difference between Example 8 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 5 mg / mL of ascorbic acid.

[0096] Example 9 The difference between Example 9 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 1 mg / mL of ascorbic acid.

[0097] Example 10 The difference between Example 10 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 10 mg / mL of sodium ascorbate and 1 mg / mL of ascorbic acid.

[0098] Example 11 The difference between Example 11 and Example 4 is only that in 6) purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, and the volume ratio of ethanol to water is 0.6:1.

[0099] Example 12 The difference between Example 12 and Example 4 is only that in the purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of genisteic acid.

[0100] Example 13 The difference between Example 13 and Example 4 is only that in the purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 1 mg / mL of genisteic acid.

[0101] Example 14 The difference between Example 14 and Example 4 is only that in the purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of L-glutathione.

[0102] Example 15 The difference between Example 15 and Example 4 is only that in the purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 0.6:1, and the mobile phase further contains 1 mg / mL of thiourea and 5 mg / mL of sodium ascorbate.

[0103] Example 16 The difference between Example 16 and Example 4 is only that in the purification by high-performance liquid chromatography, the mobile phase is a mixed system of ethanol and water, the volume ratio of ethanol to water is 1:9, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of vitamin C.

[0104] Comparative Example 1 The difference between Comparative Example 1 and Example 4 is only that in the purification by high performance liquid chromatography, the mobile phase is a mixed system of acetonitrile and water, the volume ratio of acetonitrile to water is 0.6:1, and the mobile phase further contains 5 mg / mL of sodium ascorbate and 1 mg / mL of ascorbic acid.

[0105] Comparative Example 2 The difference between Comparative Example 2 and Example 4 is only that in the purification by high performance liquid chromatography, the mobile phase is pure ethanol and it does not contain ascorbic acid, sodium ascorbate or gentisic acid.

[0106] Please refer to Table 2 for the parameters of Examples 1 to 16 and Comparative Examples 1 to 2.

[0107]

Table 2-1

[0108]

Table 2-2

[0109] As shown in Table 2 18 The initial activity data of F refers to the data that can be detected immediately after production. Usually, 18 Those skilled in the art can understand that the initial activity of F changes depending on the storage time and usage conditions. Therefore, the 18 Initial activity data of F in Examples 1 to 16 and Comparative Examples 1 to 2 are usually within 18 ±10% of the initial activity data of the target F and are within the range recognized by those skilled in the art. For example, if 10 Ci is the target 18 Initial activity of F, in actual detection, the initial activity can be 9 Ci to 11 Ci. If 6 Ci is the target 18 Initial activity of F, in actual detection, the initial activity can be 5.4 Ci to 6.6 Ci. If 3.5 Ci is the target 18 Initial activity of F, in actual detection, the initial activity can be 3.15 Ci to 3.85 Ci.

[0110] Example 17 Liquid Composition of Compound I Using the Compound I product of Example 1, a liquid composition of Compound I was formulated. The compound I was collected in a transfer bottle (i.e., prescription bottle P1) with a prescription substrate solution added in advance (for the formulation components and preparation of prescription bottle P1, refer to the preparation of prescription bottle P1. Here, the prescription substrate solution is as follows: First, dissolve another prescription bottle P2 to a predetermined volume with water, take out a part of the reagent from the pre-treated other prescription bottle P2, and mix a part of the reagent in this other prescription bottle P2 with the reagent in prescription bottle P1 to form the prescription substrate solution). Then, vitamin C and sodium ascorbate remaining in the other prescription bottle P2 (i.e., prescription bottle P2; for the formulation components and preparation of prescription bottle P2, refer to the preparation of prescription bottle P2) were added to prescription bottle P1. The activity concentration of the purified product of Compound I after mixing was 2000 MBq / mL, i.e., 54 mCi / mL. The above liquid composition was mixed with the HPLC main peak mobile phase to complete the formulation. Specifically, the formulation contains polyethylene glycol 400 0.1 g / mL, vitamin C 2 mg / mL (where the vitamin C concentration (mg / mL) / Compound I activity concentration (mCi / mL) is 0.04), sodium ascorbate 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / Compound I activity concentration (mCi / mL) is 0.37) (the mass ratio of the above vitamin C to sodium ascorbate is 1:10), water 0.816 mL / mL, and ethanol 0.094 mL / mL.

[0111] Example 18 The difference between Example 18 and Example 17 is that the formulation contains polyethylene glycol 400 0.1 g / mL, vitamin C 6 mg / mL (where the vitamin C concentration (mg / mL) / Compound I activity concentration (mCi / mL) is 0.11), sodium ascorbate 15 mg / mL (where the sodium ascorbate concentration (mg / mL) / Compound I activity concentration (mCi / mL) is 0.28) (the mass ratio of the above vitamin C to sodium ascorbate is 2:5), water 0.816 mL / mL, and ethanol 0.094 mL / mL.

[0112] Example 19 The difference between Example 19 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, vitamin C at 2 mg / mL (where the vitamin C concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.04), sodium ascorbate at 40 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.74) (the mass ratio of the vitamin C to the sodium ascorbate is 1:20), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0113] Example 20 The difference between Example 20 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, vitamin C at 8 mg / mL (where the vitamin C concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.15), sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.37) (the mass ratio of the vitamin C to the sodium ascorbate is 2:5), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0114] Example 21 The difference between Example 21 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, vitamin C at 16 mg / mL (where the vitamin C concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.3), sodium ascorbate at 40 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.74) (the mass ratio of the vitamin C to the sodium ascorbate is 2:5), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0115] Example 22 The difference between Example 22 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, gentisic acid at 2 mg / mL (where the gentisic acid concentration (mg / mL) / activity concentration of Compound I (mCi / mL) is 0.04), sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / activity concentration of Compound I (mCi / mL) is 0.37) (the mass ratio of the gentisic acid to the sodium ascorbate is 1:10), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0116] Comparative Example 3 The difference between Comparative Example 3 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0117] Comparative Example 4 The difference between Comparative Example 4 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / activity concentration of Compound I (mCi / mL) is 0.37), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0118] Comparative Example 5 The difference between Comparative Example 5 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, vitamin C at 2 mg / mL (where the vitamin C concentration (mg / mL) / activity concentration of Compound I (mCi / mL) is 0.04), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0119] Comparative Example 6 The difference between Comparative Example 6 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, L-glutathione at 2 mg / mL (where the L-glutathione concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.04), sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.37) (the mass ratio of the L-glutathione to the sodium ascorbate is 1:10), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0120] Comparative Example 7 The difference between Comparative Example 7 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, thiourea at 2 mg / mL (where the thiourea concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.04), sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.37) (the mass ratio of the thiourea to the sodium ascorbate is 1:10), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0121] Comparative Example 8 The difference between Comparative Example 8 and Example 17 is that the formulation contains polyethylene glycol 400 at 0.1 g / mL, sodium metabisulfite at 2 mg / mL (where the sodium metabisulfite concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.04), sodium ascorbate at 20 mg / mL (where the sodium ascorbate concentration (mg / mL) / the activity concentration of Compound I (mCi / mL) is 0.37) (the mass ratio of the sodium metabisulfite to the sodium ascorbate is 1:10), water at 0.816 mL / mL, and ethanol at 0.094 mL / mL.

[0122] The parameters of Examples 17 to 22 and Comparative Examples 3 to 8 are shown in Table 3.

[0123]

Table 3-1

[0124]

Table 3-2

[0125] The activity concentration of the product in Table 3 refers to the activity concentration of the purified product of Compound I, where 1 mCi = 37 MBq. The activity concentration of the purified product of Compound I refers to the activity of the Compound I product divided by the total volume of the Compound I liquid composition.

[0126] In Example 17, the activity concentration of the product was calculated to be 2000 MBq / mL, i.e., 54 mCi / mL. The ratio of the vitamin C dosage (mg / mL) to the activity concentration of Compound I (mCi / mL) was 2 to 54, i.e., 0.04, and the ratio of the sodium ascorbate dosage (mg / mL) to the activity concentration of Compound I (mCi / mL) was 20 to 54, i.e., 0.37.

[0127] Experimental Example The method for measuring the labeling rate is as follows: after the labeling reaction is completed, the sample is injected and analyzed by high-performance liquid chromatography (HPLC), and the ratio of the radioactive peak area of the target product in the liquid chromatogram to the peak areas of all radioactive peaks is calculated.

[0128] The non-decay corrected yield is determined as the ratio of the activity of the final liquid composition product measured using an activity meter to 18 the initial activity of F.

[0129] As a method for determining the radiochemical purity at 0 h, the sample is injected and analyzed by HPLC, and it is determined as the ratio of the radioactive peak area of the product to the peak areas of all radioactive peaks.

[0130] As a method for measuring the 6-h stability (radiochemical purity index), after the final product is placed at room temperature for 6 hours, the sample is injected and analyzed by HPLC, and it is determined as the ratio of the radioactive peak area of the product to the peak areas of all radioactive peaks.

[0131] The experimental effect data of Examples 1 to 16 and Comparative Examples 1 to 2 are shown in Table 4.

[0132]

Table 4

[0133] As can be seen from Table 4, in Examples 1 to 4, after the optimization of the purification process by high performance liquid chromatography, the amount of the crude product containing Compound I that can be processed increases, and thus the reaction amount of the initial activity of Compound I precursor and 18 F can be increased, and it becomes possible to provide a high-quality liquid composition of Compound I in a large batch.

[0134] Compared with Example 1, 18 when the initial activity of F is constant, in Example 2, the dosage of Compound I precursor increases, 18 the conversion of F is promoted, and the yield and labeling rate increase.

[0135] In Examples 2 to 4, when the dosage of the precursor of Compound I is constant, 18 even if the initial activity of F increases, the yield and labeling rate hardly change, and the radiochemical purity hardly changes.

[0136] Generally, when the dosage of Compound I precursor is constant, 18 the higher the initial activity of F within a certain range, the lower the yield. However, when the dosage of Compound I precursor reaches a certain amount, 18 the yield no longer changes with the change of the initial activity of F.

[0137] In Examples 4 to 6, the influence of the volume ratio of ethanol to water in the mobile phase on the final efficiency was investigated. The more ethanol in the mobile phase, the earlier the final product can be obtained. Therefore, the total preparation time is shortened and the yield is increased. Therefore, the yields of Examples 5 and 6 are slightly higher than that of Example 4. However, excessive ethanol content reduces the radiochemical purity of the final product. Therefore, the radiochemical purity of Example 5 is lower than that of Examples 4 and 6.

[0138] In Examples 4, 7, and 8, since vitamin C is not present in the mobile phase of Example 7, the yield and radiochemical purity of Example 7 are lower than those of Examples 4 and 8. Since the vitamin C content of Example 8 is increased compared to the vitamin C content of Example 4, the yield of Example 8 is superior to the yield of Example 4.

[0139] Among Examples 4 and 9 to 11, compared with Example 4, since the mobile phase of Example 9 does not contain sodium vitamin C, the yield and radiochemical purity of Example 9 are lower than those of Example 4. Since Example 10 has a higher sodium vitamin C content than Example 4, the yield of Example 10 is superior to the yield of Example 4. Since Example 11 does not contain sodium vitamin C and vitamin C, both the yield, labeling rate, and radiochemical purity decrease.

[0140] In Examples 12 and 13, compared with Example 4, in Example 12, vitamin C was changed to gentiopic acid, and the labeling rate, yield, and radiochemical purity are equivalent, indicating that the effect of gentiopic acid in the mobile phase is equivalent to that of vitamin C. Compared with Example 12, since the mobile phase of Example 13 does not contain sodium vitamin C, the yield, labeling rate, and radiochemical purity of Example 13 are all lower than those of Example 12.

[0141] In Examples 14 and 15, vitamin C was changed to other substances, and compared with Example 4, the labeling rate, yield, and radiochemical purity all decreased, indicating that vitamin C is more suitable for use in the mobile phase than L - glutathione and thiourea.

[0142] In Example 16, the ratio of ethanol to water in the mobile phase was 1:9, and as a result, the peak elution time became longer and the yield decreased.

[0143] In Comparative Example 1, the mobile phase was a mixed system of acetonitrile and water, and the use of a C18 cartridge was required in the subsequent purification step, which resulted in severe radiolysis, a longer reaction time, and a decrease in the final yield and radiochemical purity.

[0144] In Comparative Example 2, since only ethanol was used as the mobile phase, impurities and the final product came out together, and sodium ascorbate and ascorbic acid were insoluble in ethanol, so sodium ascorbate and ascorbic acid could not be contained in the mobile phase. As a result, the final Compound I product had low purity and was unstable.

[0145] The experimental effect data of Examples 17 - 22 and Comparative Examples 3 - 8 are shown in Table 5.

[0146]

Table 5

[0147] In Examples 17 - 22, the activity concentration of the product was 2000 MBq / mL, that is, 54 mCi / mL. The radiochemical purity of Examples 17 and 18 was high, and the stability for 6 hours was good.

[0148] In Example 19, the mass ratio of ascorbic acid to sodium ascorbate was 1:20, the radiochemical purity was high, the stability effect for 6 hours was good, and the sodium content was relatively high.

[0149] In Examples 20 and 21, the ratio of ascorbic acid to sodium ascorbate was 2:5, the radiochemical purity was high, and the stability effect for 6 hours was good. In Example 21, the ascorbic acid content was higher, and as a result, the pH of the liquid composition of Compound I was lower.

[0150] In Example 22, when ascorbic acid was changed to gentisic acid, the radiochemical purity was high and the stability effect for 6 hours was also good, indicating that gentisic acid in the composition can play a role equivalent to that of ascorbic acid.

[0151] From Comparative Examples 3 to 8, when the liquid composition of Compound I does not contain vitamin C or sodium ascorbate, or when vitamin C is replaced with L-glutathione or thiourea or sodium metabisulfite, the radiochemical purity at 0 hours is low, and after 6 hours, the stability effect further deteriorates, and the radiochemical purity is less than 90%, which is unqualified for myocardial perfusion PET imaging agents.

[0152] This application provides a reagent kit for the automatic preparation of the liquid composition of Compound I.

[0153] In some embodiments of this application, the configuration of the reaction bottle R1 is as shown in Table 6.

[0154]

Table 6

[0155] Preparation of potassium carbonate solution: Weigh an appropriate amount of water for injection into a glass beaker, add the prescribed amount of potassium carbonate, wash the container with a small amount of water 2 to 3 times, add it to the beaker, stir, and dissolve. Preparation of eluent: Add an appropriate amount of acetonitrile to an Erlenmeyer flask, add the prescribed amount of aminopolyether, wash the container with a small amount of acetonitrile 2 to 3 times, then add it to the Erlenmeyer flask, stir, and dissolve. Add the potassium carbonate solution prepared above, wash the beaker with the remaining amount of water in the prescribed amount 2 to 3 times, add it to the Erlenmeyer flask, and stir evenly. Finally, make up to volume with acetonitrile (add to the predetermined amount) and stir evenly. Filtration: Pass the above eluent through two 0.22 μm sterile-grade hydrophobic polytetrafluoroethylene capsule filters for filtration. Filling, sealing, capping: Dispense the filtered solution into 1 mL / bottles, stopper, and cap. Visual inspection and packaging: After passing the visual inspection, stick labels, and at the same time, use cartons to pack 1 piece / box, stick labels, seal, and package.

[0156] In some embodiments of the present application, the configuration of the reaction bottle R2 is as shown in Table 7.

[0157]

Table 7

[0158] Preparation of the compound I precursor solution: Weigh the prescribed amount of the compound I precursor acetonitrile solution into a beaker, add it to a wide-mouth bottle with a blue cap, wash the beaker 2 - 3 times with a small amount of acetonitrile, combine them and put them into the wide-mouth bottle with a blue cap, add acetonitrile to the prescribed amount (make up to volume), and stir evenly. Filtration: Filter the above-mentioned compound I precursor liquid through two 0.22 μm sterile-grade hydrophobic polytetrafluoroethylene capsule filters. Filling, sealing, capping: Dispense the filtered solution into 2 mL / bottles, plug and cap. Visual inspection and packaging: After passing the visual inspection, stick labels, and at the same time, use cartons to pack 1 piece / box, stick labels, seal and package.

[0159] In some embodiments of the present application, the configuration of the reaction bottle R3 is as shown in Table 8.

[0160]

Table 8

[0161] Preparation of the liquid: Weigh the prescribed amount of absolute ethanol, add it to the liquid preparation tank, turn on the intermediate layer cooling water, make the temperature in the tank less than 20°C, and stir evenly. Filtration: Pre-filter the solution in the preparation tank through a one-stage 5-inch 0.45 μm hydrophobic PTFE filter element, filter it through a two-stage 5-inch 0.22 μm sterile-grade hydrophobic PTFE filter element, and then push it into the receiving tank. Filling, sealing, capping: Dispense the filtered solution into 10 mL / bottles, press the plug and cap. Light inspection and packaging: After passing the light inspection, attach a label and at the same time pack it in a carton, 1 piece / box, attach a label, seal and package it.

[0162] In some embodiments of the present application, the composition of the prescription bottle P1 is as shown in Table 9.

[0163]

Table 9

[0164] Preparation of polyethylene glycol 400 solution: Weigh the prescribed amount of polyethylene glycol 400 (for injection), add an appropriate amount of injection water (<30°C) to the solution preparation barrel of polyethylene glycol 400, stir to dissolve, and store for later use. Preparation of the prescription matrix solution: Add an appropriate amount of water to the preparation tank, transfer the polyethylene glycol 400 solution to the preparation tank, wash the preparation barrel 3 times with an appropriate amount of injection water (<30°C), combine them and add them to the preparation tank, stir uniformly, then add injection water (<30°C) to the prescribed amount and stir uniformly. Filtration: Filter the solution through a one-stage 5-inch 0.45μm hydrophilic polyethersulfone filter element and a two-stage 5-inch 0.22μm sterilization-grade hydrophilic polyethersulfone filter element, and push it into the receiving tank. Filling, sealing, capping: Dispense the filtered solution into 13.8 mL / bottle, press the stopper to cover the bottle. Sterilization: After covering the bottle, sterilize the intermediate product. The sterilization conditions are a sterilization temperature of 121°C, a sterilization time of 15 minutes, and F0≥12. Light inspection and packaging: After passing the light inspection, attach a label and at the same time put the product in the bottle and the plastic inner holder into a carton and pack it 1 piece / box, attach a label, seal and package it.

[0165] In the present application, the prescription bottle P1 is used as the transfer bottle, and the reagent in the prescription bottle P1 and a part of the reagent dissolved in water in another prescription bottle P2 are combined to form the prescription matrix solution.

[0166] In some embodiments of the present application, the composition of the prescription bottle P2 is as shown in Table 10.

[0167]

Table 10

[0168] The composition of the prescription bottle P2 is as follows. Weighing: Weigh the prescribed amount of vitamin C and sodium ascorbate. Powder filling, sealing, capping: Use a high-precision packaging machine for powder filling and sealing. The filling amount of vitamin C is 1.6 g, and the filling amount of sodium ascorbate is 8.8 g. Press the stopper to cap the bottle. Light inspection and packaging: After passing the light inspection, stick a label. At the same time, put the product in the bottle and the plastic inner holder into a carton and pack them at 1 piece / box. Stick a label and seal it for packaging.

[0169] In the present application, there are two prescription bottles P2. One of the prescription bottles P2 is used for the mobile phase. The other prescription bottle P2 is used for storing the reagent after purification.

[0170] Taking Example 1 as an example, before starting the reaction, place the pre-prepared reaction bottles R1, R2, R3, and prescription bottle P1 in their corresponding positions and react them. 18 After the nucleophilic substitution reaction of F ions is completed, purify by high-performance liquid chromatography. The vitamin C and sodium ascorbate in the mobile phase are from the prescription bottle P2. Here, dissolve the other prescription bottle P2 in a predetermined volume of water in advance. Take out some reagents from the other processed prescription bottle P2. Mix a part of the reagents in this other prescription bottle P2 with the reagents in the prescription bottle P1 to form a prescription substrate solution. After purification, recover compound I into the prescription bottle P1 with the prescription substrate solution added in advance to obtain a liquid composition of compound I.

[0171] Although the present application is disclosed as above through embodiments, the above embodiments are not intended to limit the present application. Those having ordinary knowledge in the relevant technical field can make certain changes and modifications without departing from the spirit and scope of the present application. Therefore, the protection scope of the present application shall be determined by the scope of the appended patent application.

Claims

1. A method for preparing a liquid composition of Compound I, comprising the following steps: purifying the crude product containing Compound I by high-performance liquid chromatography, wherein the mobile phase used in the purification step by high-performance liquid chromatography contains ethanol and water, The compound I is 2-tert-butyl-4-chloro-5-((3-(((4-((2-(2-fluoro １８ F]ethoxy)ethoxy)methyl)-1H-1,2,3-triazol)-1-yl)methyl)benzyloxy)pyridazin-3(2H)-one, and the preparation method.

2. The preparation method according to Claim 1, wherein the mobile phase further contains one or more of sodium vitamin C, vitamin C, and gentisic acid.

3. In the purification step by high-performance liquid chromatography, in the mobile phase, the ethanol is 0.2 to 2 parts by volume with respect to 1 part by volume of water, The preparation method according to Claim 1.

4. In the mobile phase, the ethanol is 0.4 to 1 part by volume with respect to 1 part by volume of water, The preparation method according to Claim 3.

5. In the purification step by high-performance liquid chromatography, the addition amount of sodium vitamin C is 0 to 20 mg / mL, The preparation method according to Claim 2.

6. In the purification step by high-performance liquid chromatography, the addition amount of vitamin C is 0 to 10 mg / mL, The preparation method according to Claim 2.

7. In the purification step by high-performance liquid chromatography, the addition amount of gentisic acid is 0 to 10 mg / mL, The preparation method according to Claim 2.

8. The chromatography column used in the purification step by high-performance liquid chromatography is a silica gel column, The preparation method according to Claim 1.

9. Before the purification step by high-performance liquid chromatography, it further includes a step of performing a nucleophilic substitution reaction, In the nucleophilic substitution reaction, the activated １８ F ions are mixed with the precursor-containing solution of Compound I to carry out a nucleophilic substitution reaction to produce a crude product containing the Compound I, wherein the name of the precursor of Compound I is methyl 2-(2-((1-(3-((1-(tert-butyl)))-5-chloro-6-oxo-1,6-dihydropyridazin-4-yl)oxy)methyl)benzyl)-1H-1,2,3-triazol-4-yl)methoxy)ethoxy)ethyl-4-methylbenzenesulfonate, The preparation method according to Claim 1.

10. In the nucleophilic substitution reaction, the ratio of the dosage of the compound I precursor / １８ the F ion activity is in the range of (0.5 to 8):1, The unit of the dosage of the compound I precursor is mg, １８ and the unit of the F ion activity is Ci. The preparation method according to Claim 9.

11. Before the nucleophilic substitution reaction step, １８ further comprising a step of preparing F ions, The above-mentioned １８ preparation step of F ions is １８ preparation of an F ion solution, １８ concentration and elution of F ions, and １８ further includes activation of F ions. The preparation method according to Claim 9.

12. The foregoing １８ In the step of preparing the F-ion solution, １８ Transfer water containing O to the target position of the accelerator, start the accelerator to generate a proton beam, and １８ collide the water containing O, １８ generate a solution containing F ions, and the １８ initial activity of F is 0.045 Ci to 11 Ci. The preparation method according to Claim 11.

13. The above-mentioned １８ In the F-ion concentration step, The anion exchange cartridge is a tetraalkylammonium salt anion exchange cartridge, The preparation method according to Claim 11.

14. In the above-mentioned １８ in the step of eluting F ions, In the catalyst solution of cryptand and alkali metal salt, the dosage of cryptand is 5 to 40 mg, and the dosage of alkali metal salt is 1.5 to 20 mg. The preparation method according to claim 11.

15. The above-mentioned １８ In the activation step of F ions, The activation temperature is 80 to 130 °C, The temperature program control includes the following steps: 100 to 120 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation for 60 to 120 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -20 to -60 mbar, evaporation for 150 to 200 seconds; 120 to 130 °C, positive pressure 50 to 200 mbar, vacuum pressure -60 to -100 mbar, evaporation for 10 to 30 seconds; 100 to 120 °C, positive pressure 800 to 1200 mbar, vacuum pressure -800 to -1000 mbar, evaporation for 80 to 120 seconds; 80 to 100 °C, positive pressure 400 to 600 mbar, vacuum pressure -800 to -1000 mbar, evaporation for 100 to 120 seconds; 80 to 100 °C, positive pressure 600 to 900 mbar, vacuum pressure -800 to -1000 mbar, evaporation for 10 to 20 seconds. The preparation method according to claim 11.

16. Use of the liquid composition of compound I prepared by the method according to any one of claims 1 to 15 in a myocardial perfusion PET contrast agent.

17. A reagent kit for the automatic preparation of the liquid composition of compound I, comprising a reaction bottle and a formulation bottle, wherein the reaction bottle includes reaction bottle R1, reaction bottle R2, and reaction bottle R3, The reaction bottle R1 is １８ used for containing an eluent for eluting F ions, The reaction bottle R2 is １８ used to contain a solution of the compound I precursor for the nucleophilic substitution reaction of F ions, the reaction bottle R3 is used to contain reagents for diluting the crude product and flushing the reaction system, the formulation bottle includes formulation bottle P1 and formulation bottle P2, The prescription bottle P1 is, １８ used to transfer the reaction product after the nucleophilic substitution reaction of F ions, The prescription bottle P2 is １８ A reagent kit used to contain a substance used to stabilize the reaction product after the nucleophilic substitution reaction of F ions.

18. The reaction bottle R1 contains aminopolyether, potassium carbonate, water for injection, and acetonitrile. Preferably, the concentration of the aminopolyether is 5 to 40 mg / mL, the concentration of the potassium carbonate is 1.5 to 20 mg / mL, and the volume ratio of acetonitrile to water is (0.25 to 19):

1. Alternatively, the reaction bottle R2 contains １８ an acetonitrile solution of the compound I precursor for the nucleophilic substitution reaction of F ions, and preferably, the concentration of the compound I precursor in the acetonitrile solution is 1 to 10 mg / mL. Or, the reagent kit according to claim 17, wherein the reaction bottle R3 contains absolute ethanol.

19. The formulation bottle P1 contains polyethylene glycol. Preferably, the concentration of the polyethylene glycol is 0.05 to 0.3 g / mL. Or, The prescription bottle P2 is, １８ used to contain one or more of vitamin C, sodium ascorbate, and gentisic acid for stabilizing the reaction product after the nucleophilic substitution reaction of F ions. Preferably, the concentration of vitamin C is 2 to 16 mg / mL, the concentration of sodium ascorbate is 15 to 40 mg / mL, and the concentration of gentisic acid is 2 to 16 mg / mL. The reagent kit according to claim 17.

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