WO2023246830A1 - Method for preparing liquid composition of compound i and use thereof in myocardial metabolism pet imaging - Google Patents

Abstract

The present application provides a method for preparing a myocardial metabolism PET imaging agent, a liquid composition of trans-2-(2-(5-fluoro[¹⁸F]tridecyl)cyclopropyl)acetic acid (compound I for short), and use thereof, wherein a crude product containing compound I is purified by high performance liquid chromatography; wherein the mobile phase used in the high performance liquid chromatography purification step comprises ethanol and water. The present application optimizes the experimental process and adjusts the amount of compound I tert-butyl ester precursor to shorten the reaction time required to achieve the same labeling rate, and increases the radioactivity of the initial ¹⁸F ions to increase the labeling rate, thereby increasing the yield.

Description

Preparation method of a compound I liquid composition and its use in myocardial metabolism PET imaging Technical field

This application belongs to the field of chemical pharmaceutical technology, and in particular relates to a preparation method of a liquid composition of Compound I and its use in PET imaging of myocardial metabolism.

Background technique

As the most advanced imaging technology in the field of biomedical engineering today, molecular medical imaging technology uses imaging methods to conduct qualitative and quantitative research on biological processes in living bodies at the cellular and molecular levels, and to analyze the physiology and pathology of organisms at the molecular level. Real-time, dynamic, in-body, non-invasive imaging technology for changes in the body. It is the key and core technology for studying targeted and specific molecular probes and therapeutic drugs. Multimodal molecular imaging technology can realize the complementary advantages of different imaging equipment, making the imaging results obtained more accurate and reliable. Clinical practice has proven that multi-modal molecular medical imaging equipment plays an important role in early diagnosis and treatment of major diseases, formulation of treatment plans, and verification and evaluation of treatment effects.

Positron Emission Tomography (PET) equipment is mainly functional imaging. It consists of a PET imaging device and a radioactive tracer injected into the patient's blood. A frequently used radioactive tracer is Fluoro-deoxy-glucose (FDG), which is a compound synthesized from a simple sugar and a small amount of radioactive fluorine. After the human body injects [ ^18F ]FDG , it will accumulate in tissues and organs in the body. At the same time, ¹⁸ F will decay and emit a positron. The positron will undergo an annihilation reaction to produce a pair of photons moving in the opposite direction, with an energy of 511keV. PET equipment can detect these photons and record the information of this pair of photons. Through this information, the position of positron annihilation in the body can be restored, thereby obtaining the distribution map of [ ¹⁸ F]FDG in the human body.

However, there are still some limitations in the clinical application of ¹⁸ F-FDG in brain tumor imaging and differential diagnosis of tumors and inflammation, which uses glucose metabolism as an energy substrate.

Contents of the invention

This application provides a preparation method and use of Compound I that can be used for cardiac imaging.

The technical solution for this application is as follows:

1. A method for preparing a compound I liquid composition,

The crude product containing compound I is purified by high performance liquid chromatography; wherein, the mobile phase used in the high performance liquid chromatography purification step includes ethanol and water;

Compound I is trans-2-(2-(5-fluoro[ ^18F ]tridecyl)cyclopropyl)acetic acid.

2. According to the preparation method described in item 1,

In the HPLC purification step:

In the mobile phase,

Relative to 1 volume part of water, the ethanol is 2-5 parts by volume;

Preferably, the ethanol is 3-4 parts by volume relative to 1 part by volume of water.

3. According to the preparation method described in item 1,

In the HPLC purification step:

The mobile phase also includes vitamin C.

4. According to the preparation method described in item 3,

In the HPLC purification step:

In the mobile phase, the added amount of vitamin C is 0.1 mg/mL-10 mg/mL; preferably, it is 0.1 mg/mL-5 mg/mL.

5. According to the preparation method described in item 1,

In the HPLC purification step:

The mobile phase also includes gentisic acid.

6. According to the preparation method described in item 5,

In the HPLC purification step:

In the mobile phase, the added amount of gentisic acid is 0.1 mg/mL-10 mg/mL; preferably, it is 0.1 mg/mL-5 mg/mL.

7. According to the preparation method described in item 1,

The chromatographic column is a silica gel column, preferably a reversed-phase C18 silica gel column, and further preferably an XBridge BEH C18 OBD Prep column;

More preferably, isocratic elution is used, and the elution flow rate of the mobile phase is 3 mL/min-6 mL/min.

8. According to the preparation method described in item 1,

Before the high-performance liquid chromatography purification step, a nucleophilic substitution reaction step and a de-tert-butyl esterification reaction step are also included:

Nucleophilic substitution reaction: Mix the activated ¹⁸ F ions with a solution containing the precursor of Compound I tert-butyl ester to perform a nucleophilic substitution reaction to obtain an intermediate product solution containing Compound I tert-butyl ester;

De-tert-butyl esterification reaction: Add an acidic solvent to the intermediate product solution of the above-mentioned compound I tert-butyl ester to perform the de-tert-butyl esterification reaction to obtain a product containing compound I;

The precursor of compound I tert-butyl ester is trans-tert-butyl 2-(2-(5-(methanesulfonyloxy)tridecyl)cyclopropyl)acetate;

Compound I tert-butyl ester is trans-2-(2-(5-fluoro[ ^18F ]tridecyl)cyclopropyl)acetate tert-butyl ester.

9. The preparation method according to item 8,

In nucleophilic substitution reactions,

The starting activity ratio range of compound I tert-butyl ester precursor/ ¹⁸ F is (0.2-5):1;

Wherein, the weight unit of the compound I tert-butyl ester precursor is mg, and the unit of initial activity is Ci.

10. The preparation method according to item 9,

The initial activity of ¹⁸ F is 0.09Ci-11Ci, preferably 3.6Ci-11Ci.

11. The preparation method according to item 8,

In the de-tert-butyl esterification reaction,

The acidic solvent is a mixed system of trifluoroacetic acid and acetonitrile;

Preferably, trifluoroacetic acid is 0.4-2 parts by volume relative to 1 part by volume of acetonitrile;

Further preferably, the amount of trifluoroacetic acid is 0.6-1.5 parts by volume relative to 1 part by volume of acetonitrile.

12. Use of the compound I liquid composition prepared according to the method described in any one of items 1 to 11 in the preparation of a PET imaging agent for myocardial metabolism.

In addition, this application provides a liquid composition containing Compound I, the specific scheme is as follows:

1. A liquid composition comprising compound I, wherein the liquid composition further includes vitamin C and/or gentisic acid;

Compound I is trans-2-(2-(5-(fluoro[ ^18F ])tridecyl)cyclopropyl)acetic acid.

2. The liquid composition according to item 1, the vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) ratio range is (0.009-0.2):1.

3. The liquid composition according to item 2,

The ratio range of vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is (0.009-0.1):1.

4. The liquid composition according to item 1, gentisic acid concentration (mg/mL)/compound I activity concentration The degree (mCi/mL) ratio range is (0.009-0.2):1.

5. The liquid composition according to item 4,

The ratio range of gentisic acid concentration (mg/mL)/compound I activity concentration (mCi/mL) is (0.009-0.1):1.

6. The liquid composition of item 1, further comprising polysorbate 80.

7. The liquid composition according to item 6,

The polysorbate 80 concentration is 0.5mg/mL-2mg/mL.

8. A method for preparing the liquid composition according to any one of items 1 to 7, wherein,

The ethanol aqueous solution containing vitamin C and/or gentisic acid and compound I is collected from the purification step to obtain a liquid composition containing compound I.

9. According to the method described in item 8,

Mix vitamin C and/or gentisic acid, sodium chloride, and polysorbate 80 with an ethanol aqueous solution containing vitamin C and/or gentisic acid and compound I to obtain a liquid composition containing compound I.

10. Use of the compound I liquid composition according to any one of items 1 to 7, or the compound I liquid composition prepared by the method described in item 8 or 9, in the preparation of a PET imaging agent for myocardial metabolism.

Compared with the existing technology, the beneficial effects of this application are:

(1) This application optimizes the experimental process plan, changes the dosage of the tert-butyl ester precursor of Compound I, shortens the reaction time to achieve the same labeling rate, increases the radioactive activity of the initial ¹⁸ F ions, and increases the labeling rate, thereby improving Yield. The process parameters and process flow are clear and specific, which can be applied to large-volume activity production and meet the automation needs.

(2) In the optimized technical solution of this application, an anti-radiation decomposition agent is used in the mobile phase of the purification process to avoid product loss due to radiation decomposition during the purification and formulation process, thereby increasing the yield. In addition, this application uses an anti-radiation decomposition agent during the formulation process to ensure the stability of the product.

(3) This application optimizes the purification process, removes the C18 column, and uses ethanol/water system instead of acetonitrile/water system as the mobile phase. This simplifies time and improves the radiochemical purity and stability of the product.

Detailed ways

Compound Ⅰ, the chemical name is trans-2-(2-(5-(fluoro[ ¹⁸ F]) tridecyl) cyclopropyl) acetic acid, and the chemical structural formula is:

The chemical formula is: C ₁₈ H ₃₃ ¹⁸ FO ₂

Molecular weight: 299.46

In this application, ¹⁸ F-labeled myocardial metabolism PET imaging agent refers to the compound I liquid composition.

Compound I tert-butyl ester precursor, the chemical name is trans-tert-butyl 2-(2-(5-(methanesulfonyloxy)tridecyl)cyclopropyl)acetate, and the chemical structural formula is:

The chemical formula is: C ₂₃ H ₄₄ O ₅ S

Molecular weight is: 432.66

Compound Ⅰ tert-butyl ester, the chemical name is trans-2-(2-(5-fluoro[ ^18F ]tridecyl)cyclopropyl)acetate tert-butyl ester, and the chemical structural formula is:

The chemical formula is: C ₂₂ H ₄₁ ¹⁸ FO ₂

Molecular weight: 355.57

Aminopolyether (K ₂₂₂ ) is a three-bridged crown ether molecule with a cryptoid cavity. It is a typical aza cryptether and a kind of cryptether. Due to its unique coordination characteristics, azacryptoethers can well select cations to complex transition metals and heavy metals. The resulting complexes are more stable and are lipophilic and hydrophilic, so they have good research potential. prospect.

In the existing technology, the classic synthesis method of aminopolyether (K ₂₂₂ ) is the highly dilution method proposed by Lehn et al., which is one of the typical non-template ion synthesis methods. The specific steps are as follows: the raw material 1,8-diamino-3 , 6-dioxaoctane and 1,8-dioxanoyl chloride-3,6-dioxaoctane were dissolved in a large amount of benzene solvent and heated for 8 hours, then reduced by lithium aluminum tetrahydride for 24 hours, and then passed through Column chromatography separation and recrystallization gave aminopolyether (K ₂₂₂ ). This method requires a large amount of solvent, such as benzene, has a long synthesis route, complicated operations, low yield, and low economic benefits. In addition to the high dilution method, another classic synthesis method of aminopolyether (K ₂₂₂ ) The method proposed by Kulstad and Malmsten uses Na ₂ CO ₃ as a template to obtain the sodium iodide complex of the aminopolyether (K ₂₂₂ ) in acetonitrile, and then decomplexes it through the resin to obtain the aminopolyether (K ₂₂₂ ). resolve resolution. The specific steps are as follows: react the raw materials 1,2-bis(2-iodoethoxy)ethane and benzylamine in an acetonitrile solution under reflux for 3 days, and then obtain an intermediate through post-processing. The intermediate is recrystallized with acetone. After filtration, a NaI complex is obtained. The complex is decomplexed through cation exchange resin and anion exchange resin under acidic conditions to prepare amino polyether (K ₂₂₂ ). This method has simple equipment, less solvent consumption, and relatively mild reaction conditions. However, the applicant found through research that when the content of sodium ions is reduced to a certain amount using the method of decomplexing through ion exchange resin, the decomplexation cannot proceed and the yield is low.

This application provides a method for preparing compound I. The synthesis route is as follows:

This application provides a preparation method of compound I, wherein,

The crude product containing compound I is purified by high performance liquid chromatography;

Wherein, the mobile phase used in the high performance liquid chromatography purification step includes ethanol and water.

In some embodiments of the present application, in the high performance liquid chromatography purification step: in the mobile phase, the ethanol is 2-5 parts by volume relative to 1 part by volume of water; preferably, relative to 1 part by volume parts of water, and the ethanol is 3-4 parts by volume;

For example, relative to 1 part by volume of water, the ethanol may be 2 parts by volume, 3 parts by volume, 4 parts by volume, 5 parts by volume, or any range therebetween.

In some embodiments of the present application, in the high performance liquid chromatography purification step: the mobile phase further includes vitamin C.

In some embodiments of the present application, in the mobile phase, the added amount of vitamin C is 0.1 mg/mL-10 mg/mL; preferably 0.1 mg/mL-5 mg/mL;

For example, the vitamin C addition amount can be 0.1 mg/mL, 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 , 10mg/mL or any range in between.

In some embodiments of the present application, in the high performance liquid chromatography purification step: the mobile phase Gentisic acid is also included.

In some embodiments of the present application, in the mobile phase, the added amount of gentisic acid is 0.1 mg/mL-10 mg/mL; preferably 0.1 mg/mL-5 mg/mL;

For example, the added amount of gentisic acid can be 0.1 mg/mL, 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. mL, 10 mg/mL, or any range in between.

In some embodiments of the present application, the mobile phase may contain both vitamin C and gentisic acid, or may contain only one of vitamin C and gentisic acid.

In some embodiments of the present application, in the high-performance liquid chromatography purification step, the chromatographic column is a silica gel column, preferably a reverse-phase C18 silica gel chromatographic column, and further preferably an XBridge BEH C18 OBD Prep column.

In some embodiments of the present application, isocratic elution is used, and the elution flow rate of the mobile phase is 3mL/min-6mL/min;

For example, the elution flow rate of the mobile phase may be 3 mL/min, 4 mL/min, 5 mL/min, 6 mL/min or any range therebetween.

In some embodiments of the present application, before the high-performance liquid chromatography purification step, a nucleophilic substitution reaction step and a de-tert-butyl esterification reaction step are also included:

Nucleophilic substitution reaction: Mix the activated ¹⁸ F ions with a solution containing the precursor of Compound I tert-butyl ester to perform a nucleophilic substitution reaction to obtain an intermediate product solution containing Compound I tert-butyl ester;

De-tert-butyl esterification reaction: Add an acidic solvent to the intermediate product solution of the above compound I tert-butyl ester to perform the de-tert-butyl esterification reaction to obtain a product containing compound I.

In some embodiments of the present application, in a nucleophilic substitution reaction,

The starting activity ratio range of compound I tert-butyl ester precursor/ ¹⁸ F is (0.2-5):1;

Wherein, the weight unit of the compound I tert-butyl ester precursor is mg, and the unit of initial activity is Ci;

For example, the starting activity ratio of Compound I tert-butyl ester precursor/ ^18F can be 0.2:1, 0.3:1, 0.5:1, 0.7:1, 1:1, 2:1, 3:1, 4:1 , 5:1 or any range in between.

In some embodiments of the present application, the initial activity of ¹⁸ F is 0.09Ci-11Ci, preferably 3.6Ci-11Ci;

For example, the ¹⁸ F initial activity is 0.09Ci, 0.1Ci, 0.5Ci, 1Ci, 2Ci, 3Ci, 4Ci, 5Ci, 6Ci, 7Ci, 8Ci, 9Ci, 10Ci, 11Ci or any range therebetween.

In some embodiments of the present application, in the de-tert-butyl esterification reaction, the acidic solvent is tris Fluoroacetic acid and acetonitrile; relative to 1 part by volume of acetonitrile, trifluoroacetic acid is 0.4-2 parts by volume; further preferably, relative to 1 part by volume of acetonitrile, trifluoroacetic acid is 0.6-1.5 parts by volume;

For example, relative to 1 part by volume of acetonitrile, trifluoroacetic acid is 0.4 parts by volume, 0.5 parts by volume, 0.6 parts by volume, 0.7 parts by volume, 0.8 parts by volume, 0.9 parts by volume, 1 part by volume, 1.5 parts by volume, 2 parts by volume, or any range in between.

The present application also provides a liquid composition containing compound I, wherein the liquid composition further includes vitamin C and/or gentisic acid.

In some embodiments of the present application, the ratio range of vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is (0.009-0.2):1, preferably (0.009-0.1):1;

For example, the ratio of vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) can be 0.009:1, 0.01:1, 0.02:1, 0.03:1, 0.04:1, 0.05:1, 0.06: 1. 0.07:1, 0.08:1, 0.09:1, 0.1:1, 0.11:1, 0.12:1, 0.13:1, 0.14:1, 0.15:1, 0.16:1, 0.17:1, 0.18:1, 0.19:1, 0.2:1, or any range in between.

In some embodiments of the present application, the ratio range of gentisic acid concentration (mg/mL)/compound I activity concentration (mCi/mL) is (0.009-0.2):1, preferably (0.009-0.1):1;

For example, the ratio of gentisic acid concentration (mg/mL)/compound I activity concentration (mCi/mL) can be 0.009:1, 0.01:1, 0.02:1, 0.03:1, 0.04:1, 0.05:1, 0.06 :1, 0.07:1, 0.08:1, 0.09:1, 0.1:1, 0.11:1, 0.12:1, 0.13:1, 0.14:1, 0.15:1, 0.16:1, 0.17:1, 0.18:1 , 0.19:1, 0.2:1 or any range in between.

In some embodiments of the present application, the liquid composition containing compound I may contain both vitamin C and gentisic acid, or may contain only one of vitamin C and gentisic acid.

In some embodiments of the present application, the liquid composition further includes polysorbate 80.

In some embodiments of the present application, the polysorbate 80 concentration is 0.5 mg/mL-2 mg/mL;

For example, the polysorbate 80 concentration may be 0.5 mg/mL, 0.8 mg/mL, 1 mg/mL, 1.5 mg/mL, 2 mg/mL or any range therebetween.

The present application also provides a method for preparing the above liquid composition, wherein:

The ethanol aqueous solution containing vitamin C and/or gentisic acid and compound I is collected from the purification step to obtain a liquid composition containing compound I.

In some embodiments of the present application, vitamin C and/or gentisic acid, sodium chloride, polysan Pear ester 80 is mixed with an ethanol aqueous solution containing vitamin C and/or gentisic acid and compound I to obtain a liquid composition containing compound I.

This application also provides the use of the above-mentioned Compound I liquid composition and the Compound I liquid composition prepared by the above-mentioned method as a myocardial metabolism PET imaging agent.

In this application, ^{the 18} F initial activity, also known as ^{the 18} F ion activity, refers to the production of a solution containing ¹⁸ F ions after starting the accelerator to produce a proton beam that bombards oxygen-containing [ ¹⁸ O] water. ^{The 18} F ion activity measured by a meter.

In this application, ^{the 18} F initial activity refers to the concentration that can be detected after producing a solution containing ¹⁸ F ions after bombarding oxygen-containing [ ¹⁸ O] water with a proton beam generated by a startup accelerator. Detectable refers to A reasonable detection time can be controlled by those skilled in the art, such as within 10 minutes after production. In addition, those skilled in the art can understand that as the storage time after production changes, the initial activity of ¹⁸ F will change to a certain extent, but usually the error range is within the range of ±10%.

In some embodiments of the present application, the initial activity of ¹⁸ F is 0.09Ci-11Ci; preferably 3.6Ci-11Ci; for example, the initial activity of ^18F can be 0.09Ci, 0.1Ci, 0.5Ci, 1Ci , 2Ci, 3Ci, 4Ci, 5Ci, 6Ci, 8Ci, 9Ci, 10Ci, 11Ci or any range in between.

In this application, large batch refers to products with high total activity, which generally refers to products with total activity exceeding 1 Ci or 37GBq;

High activity concentration products generally refer to products with an activity concentration exceeding 50mCi/mL, that is, 1850MBq/mL.

In this application, the labeling rate refers to the labeling reaction between ^18F and the reaction precursor. ^18F replaces the leaving group in the precursor and converts it into the final labeled product. The labeled product contains ^18F , so the labeling rate is defined as The activity of the labeled product is higher than the total ¹⁸ F activity participating in the reaction.

In this application, the yield refers to the ratio of the activity of the final product Compound I liquid composition to the starting activity of ¹⁸ F.

In some embodiments of the present application, before the nucleophilic substitution reaction step, a ¹⁸ F ion preparation step is also included; the ¹⁸ F ion preparation step also includes ¹⁸ F ion solution preparation, ¹⁸ F ion enrichment and elution, ¹⁸ F ion activation;

Among them, ¹⁸ F ion solution preparation: the accelerator prepares ¹⁸ F ion solution;

¹⁸ F ion enrichment: The ¹⁸ F ion solution prepared above is enriched through an anion exchange column;

¹⁸ F ion elution: use cryptether and alkali metal salt catalyst solution to elute ¹⁸ F ions;

¹⁸ F ion activation: The solvent is blown dry by programmed control of temperature, nitrogen or other inert gas, and the ¹⁸ F ions are activated to obtain activated ¹⁸ F ions.

In some embodiments of the present application, in the ¹⁸ F ion solution preparation step, water containing ¹⁸ O is transported to the accelerator target position, and the accelerator is started to generate a proton beam to bombard the water containing ¹⁸ O to produce a solution containing ¹⁸ F ions. .

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

In some embodiments of the present application, in the ¹⁸ F ion elution step, in the cryptether and alkali metal salt catalyst solution, the dosage of cryptether is 5 mg-40 mg, and the dosage of alkali metal salt is 1.5 mg-20 mg; in the solution The dosage of cryptether can be 5mg, 8mg, 10mg, 15mg, 20mg, 40mg or any range between them; the dosage of alkali metal salt in the solution can be 1.5mg, 3mg, 5mg, 10mg, 20mg or any range between them.

In some embodiments of the present application, the cryptether is 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo[8,8,8]hexadecane ( Kryptofix-2.2.2, aminopolyether); the alkali metal salt is one or more of K ₂ CO ₃ , Na ₂ CO ₃ , Cs ₂ CO ₃ , KHCO ₃ and NaHCO ₃ .

In some embodiments of the present application, the catalyst solution is selected from a mixed solvent system of acetonitrile and water, wherein the volume ratio of acetonitrile to water is (0.2-10):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 any range in between; the volume of acetonitrile and water mixed solvent is 0.3mL-2mL.

In some embodiments of the present application, in the ¹⁸ F ion activation step, the activation temperature is 80°C-130°C;

Among them, the program control temperature includes the following steps: 100-120°C, positive pressure 50-200mbar, vacuum pressure -20~-60mbar, evaporation 60-120s; 120-130°C, positive pressure 50-200mbar, vacuum pressure -20~- 60mbar, evaporation 150-200s; 120-130℃, positive pressure 50-200mbar, vacuum pressure -60～-100mbar, evaporation 10-30s; 100-120℃, positive pressure 800-1200mbar, vacuum pressure -800～-1000mbar, Evaporation 80-120s; 80-100℃, positive pressure 400-600mbar, vacuum pressure -800～-1000mbar, evaporation 100-120s; 80-100℃, positive pressure 600-900mbar, vacuum pressure -800～-1000mbar, evaporation 10 -20s.

Reactions with protective groups (such as tert-butyl ester group protecting carboxyl group, tert-butoxycarbonyl group protecting amino group, etc.) are all carried out using a two-step method.

In this application, the dosage of fluorine [ ¹⁸ F] ions is increased from 4Ci level to 10Ci level. The preparation method of the present application can be applied to higher ¹⁸ F ion starting amounts.

The preparation method of the above-mentioned compound I provided by this application can increase the yield to more than 30% and shorten the production time from about 80 minutes to about 60 minutes. In this preparation method, since the solvent used in the de-tert-butyl esterification reaction is acetonitrile, the step of nitrogen removal of the solvent can be omitted and the process can be shortened. The use of trifluoroacetic acid/acetonitrile system in the de-tert-butyl esterification reaction reduces the risk of solvent residues in the final product, while increasing the initial activity of compound I tert-butyl ester precursor and ¹⁸ F, and improving the conversion of this step. efficiency; the mobile phase ethanol/water mixed solvent system used in the high-performance liquid chromatography purification step omits the subsequent C18 column enrichment purification and absolute ethanol elution steps, avoiding the radiation of the product under high activity Decompose and shorten the preparation process. In addition, the anti-radiation decomposition agent vitamin C is added to the mobile phase to avoid radiation decomposition during the column purification process.

This application provides a general and/or specific description of the materials and test methods used in the test. In the following examples, if there are no other special instructions, % means wt%, that is, weight percentage. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional reagent products that are commercially available. Table 1 shows the sources of raw materials used in the examples.

The parameters set in this embodiment are preferred embodiments of this application, and those skilled in the art can predict that other parameter ranges will also achieve the technical effects described in this application.

Sources of raw materials used in the examples in Table 1

Example

The automation equipment is AllinOne equipment from Trasis Company. The power unit of the equipment 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 automated equipment, which is placed in a radiation shielding box, it can protect operators from radiation damage and increase the operating dose. At the same time, due to computer control, the process steps can be controlled more accurately and with higher repeatability. Reduce human bias.

Example 1 Preparation of compound I

1) Preparation of ¹⁸ F ion solution

Transfer 2g of water containing oxygen [ ¹⁸ O] to the accelerator target position, start the accelerator to generate a proton beam to bombard the water containing oxygen [ ¹⁸ O], and produce a solution containing ¹⁸ F ions, with an initial activity of ¹⁸ F 4Ci.

2) ¹⁸ F ion enrichment

Pass the solution of ¹⁸ F ions prepared above through an anion exchange solid phase extraction cartridge (Waters brand QMA cartridge. The QMA cartridge is preferably washed with 5 mL of 0.5 mol/L K ₂ CO ₃ and then with 10 mL of water. After washing and activation, ¹⁸ F ions are enriched on the QMA cartridge.

3) ¹⁸ F ion elution

Use cryptether and alkali metal salt catalyst solution to elute and elute ¹⁸ F ions into the reaction bottle. Specifically, mix K ₂₂₂ 8 mg (dissolved in 0.3 mL acetonitrile) and K ₂ CO ₃ 4 mg (dissolved in 0.3 mL water). Prepare a mixed solution of acetonitrile and water, elute the above-mentioned QMA cartridge, and elute the K ¹⁸ F/K ₂₂₂ complex into the reaction bottle.

4) ¹⁸ F ion activation

The ¹⁸ F ions eluted in step 3) are programmed to heat at 100°C to 125°C under nitrogen flow to dry the solvent to obtain activated ¹⁸ F ions.

5) ¹⁸ F ion nucleophilic substitution reaction

Add 1 mL of acetonitrile solution containing compound I tert-butyl ester precursor to the reaction bottle. The dosage of compound I tert-butyl ester precursor is 3 mg. The dosage of compound I tert-butyl ester precursor (mg)/ ¹⁸ F initial activity (Ci) It is 3mg/4Ci (i.e. 0.75:1), heated to 120°C under sealed conditions, and reacted for 10 minutes. The precursor of compound I tert-butyl ester and K ¹⁸ F/K ₂₂₂ carried out nucleophilic substitution reaction to obtain compound I tert-butyl ester. intermediate products.

6) Detert-butyl esterification reaction

Add 1 mL of a mixed system of trifluoroacetic acid and acetonitrile (the volume ratio of trifluoroacetic acid and acetonitrile is 1:1) to the above intermediate product containing compound I tert-butyl ester, the reaction temperature is 70°C, the reaction time is 2 min, and the reaction time is 2 min. The tert-butyl ester protecting group was removed to obtain a crude product containing compound I. After the reaction was completed, nitrogen flow Heat to 100°C to remove the solvent.

7) Purification by high performance liquid chromatography

Load the crude product containing compound I into the injection loop and purify according to the following chromatographic conditions: HPLC eluate is obtained

Column: XBridge BEH C18 OBD Prep column, 130A, 5μm, 10×250mm

Mobile phase: a mixed solvent system of ethanol and water, where the volume ratio of ethanol to water is 3:1. The mobile phase also includes vitamin C, and the concentration of vitamin C is 0.5mg/mL;

Flow rate: 5mL/min

Detector: Radioactivity Detector

Monitor and track the radioactive signal, and collect the main radioactive peak of Compound I to the transfer bottle.

After purification, pure compound I was obtained.

Example 2

The difference between Example 2 and Example 1 is that: the dosage of Compound I tert-butyl ester precursor is 6 mg, and the dosage of Compound I tert-butyl ester precursor (mg)/ ¹⁸ F initial activity (Ci) is 6 mg/4Ci (i.e. 1.5 :1)Other conditions are the same.

Example 3

The difference between Example 3 and Example 1 is that: the dosage of Compound I tert-butyl ester precursor is 10 mg, and the dosage of Compound I tert-butyl ester precursor (mg)/ ¹⁸ F initial activity (Ci) is 10 mg/4Ci (i.e. 2.5 :1), the rest of the conditions are the same.

Example 4

The difference between Example 4 and Example 3 is that: in the preparation of ¹⁸ F ion solution, the initial activity of ¹⁸ F is 6.5Ci, and the dosage of compound I tert-butyl ester precursor (mg) / ¹⁸ F initial activity (Ci) is 10 mg /6.5Ci (ie 1.54:1), other conditions are the same.

Example 5

The difference between Example 5 and Example 3 is that: in the preparation of ¹⁸ F ion solution, the initial activity of ¹⁸ F is 10Ci, and the dosage of compound I tert-butyl ester precursor (mg) / ¹⁸ F initial activity (Ci) is 10mg/ 10Ci (i.e. 1:1), other conditions are the same.

Example 6

The difference between Example 6 and Example 5 is that in the de-tert-butyl esterification reaction, the volume ratio of trifluoroacetic acid and acetonitrile is 0.67:1, and the other conditions are the same.

Example 7

The difference between Example 7 and Example 5 is that in the de-tert-butyl esterification reaction, the volume ratio of trifluoroacetic acid and acetonitrile is 0.43:1, and the other conditions are the same.

Example 8

The difference between Example 8 and Example 5 is that during purification by high-performance liquid chromatography, the mobile phase is a mixed solvent system of ethanol and water, where the volume ratio of ethanol to water is 4:1, and the other conditions are the same.

Example 9

The difference between Example 9 and Example 5 is that during purification by high performance liquid chromatography, the mobile phase is a mixed solvent system of ethanol and water, in which the volume ratio of ethanol to water is 2:1, and the other conditions are the same.

Example 10

The difference between Example 10 and Example 5 is that during purification by high-performance liquid chromatography, the concentration of vitamin C in the mobile phase is 1 mg/mL, and the other conditions are the same.

Example 11

The difference between Example 11 and Example 5 is that during purification by high-performance liquid chromatography, the concentration of vitamin C in the mobile phase is 5 mg/mL, and the other conditions are the same.

Example 12

The difference between Example 12 and Example 5 is that during purification by high-performance liquid chromatography, the concentration of vitamin C in the mobile phase is 10 mg/mL, and the other conditions are the same.

Example 13

The difference between Example 13 and Example 5 is that during purification by high performance liquid chromatography, the mobile phase does not contain vitamin C, and the other conditions are the same.

Example 14

The difference between Example 14 and Example 5 is that during purification by high-performance liquid chromatography, the mobile phase also includes L-glutathione and does not contain vitamin C. The concentration of L-glutathione is 0.5 mg/mL, and the remaining Conditions are the same.

Example 15

The difference between Example 15 and Example 5 is that during high-performance liquid chromatography purification, the mobile phase also includes thiourea and does not contain vitamin C. The concentration of thiourea is 0.5 mg/mL, and the other conditions are the same.

Example 16

The difference between Example 16 and Example 5 is that in the de-tert-butyl esterification reaction, the volume ratio of trifluoroacetic acid and dichloromethane is 0.18:1, and the other conditions are the same.

Example 17

The difference between Example 17 and Example 5 is that during high-performance liquid chromatography purification, the mobile phase also includes gentisic acid and does not contain vitamin C. The concentration of gentisic acid is 0.5 mg/mL, and the other conditions are the same.

Example 18

The difference between Example 18 and Example 5 is that during purification by high-performance liquid chromatography, the mobile phase also includes vitamin C and gentisic acid. The concentration of vitamin C is 0.25 mg/mL, and the concentration of gentisic acid is 0.25 mg/mL. , other conditions are the same.

Comparative example 1

The difference between Comparative Example 1 and Example 5 is that during high-performance liquid chromatography purification, the mobile phase is a mixed solvent system of acetonitrile and water, where the volume ratio of acetonitrile and water is 3:1, and the mobile phase does not contain vitamin C. , other conditions are the same.

Table 2 Parameters of Examples 1-18 and Comparative Example 1

In Table 2, column 1 is XBridge BEH C18 OBD Prep column, 130A, 5μm, 10×250mm. In addition, the ^18F initial activity data shown in Table 2 refers to the data that can be detected after production, but Those skilled in the art can understand that usually the initial activity of ¹⁸ F will change with the storage time and use conditions. Therefore, the data of the initial activity of ¹⁸ F in Examples 1-18 and Comparative Example 1 are usually the target. The range of ¹⁸ F initial activity data within ±10% is within the range recognized by those skilled in the art. For example, 10Ci is the target ¹⁸ F initial activity. In actual detection, the initial activity can be 9Ci-11Ci. , 1Ci is the starting activity of ¹⁸ F of the target, in actual detection the starting activity can be 0.9Ci-1.1Ci, 4Ci is the starting activity of ¹⁸ F of the target, in actual detection the starting activity can be 3.6 Ci-4.4 Ci.

Example 21 Compound I liquid composition

Take the compound I product of Example 1, formulate a compound I liquid composition, and collect the compound I into a transfer bottle that has been added to the prescription in advance. After mixing, the activity concentration of pure compound I is 2000MBq/mL, that is, 54mCi/mL. The prescription in the transfer bottle is mixed with the HPLC main peak mobile phase to complete the prescription. Specifically, the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, and 0.5 mg/mL vitamin C (wherein , Vitamin C concentration (mg/mL)/Compound I activity concentration (mCi/mL) is 0.009).

Specifically, the above prescription component Compound I liquid composition is added to an ethanol aqueous solution containing a small amount of vitamin C and Compound I, so that the concentration of the above components meets the above conditions.

In the liquid composition of compound I, absolute ethanol and polysorbate 80 serve as co-solvents to help dissolve compound I, and vitamin C, as an anti-radiation decomposition agent, can prevent compound I from decomposing due to radiation and improve its stability.

Example 22

The difference between Example 22 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 2 mg/mL polysorbate 80, 0.5 mg/mL of vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009), and the other conditions are the same.

Example 23

The difference between Example 23 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, and 0.5 mg/mL polysorbate 80, 0.5 mg/mL vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009), and the other conditions are the same.

Example 24

The difference between Example 24 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 1 mg /mL of vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.018), and the other conditions are the same.

Example 25

The difference between Example 25 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 5 mg /mL of vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.09), and the other conditions are the same.

Example 26

The difference between Example 26 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 10 mg /mL of vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.18), and the other conditions are the same.

Example 27

The difference between Example 27 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 0.5 mg/mL of gentisic acid (wherein, gentisic acid concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Specifically, in an ethanol aqueous solution containing a small amount of gentisic acid and Compound I, the above prescription components are added to obtain a Compound I liquid composition, so that the concentration of the above components meets the above conditions.

Comparative example 2

The difference between Comparative Example 2 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, and 0.5 mg/mL vitamin C (wherein, Vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Comparative example 3

The difference between Comparative Example 3 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polyethylene glycol 400, 0.5 mg/mL vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Comparative example 4

The difference between Comparative Example 4 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL propylene glycol, 0.5 mg/mL of vitamin C (wherein, vitamin C concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Comparative example 5

The difference between Comparative Example 5 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, and 1 mg/mL polysorbate 80.

Comparative example 6

The difference between Comparative Example 6 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 0.5 mg/mL of L-glutathione (where L-glutathione concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Comparative example 7

The difference between Comparative Example 7 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 0.5 mg/mL of thiourea (wherein, thiourea concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Comparative example 8

The difference between Comparative Example 8 and Example 21 is that the compound I liquid composition includes 6.2 mg/mL sodium chloride, 0.88 mL/mL water, 0.12 mL/mL ethanol, 1 mg/mL polysorbate 80, 0.5 mg/mL sodium metabisulfite (wherein, sodium metabisulfite concentration (mg/mL)/compound I activity concentration (mCi/mL) is 0.009).

Table 3 Parameters of Examples 21-27 and Comparative Examples 2-8

Experimental example

The measurement method of labeling rate is: after the labeling reaction is completed, use high-performance liquid chromatography (HPLC) to inject and analyze the sample. The ratio of the radioactive peak area of the target product in the liquid chromatogram to the peak area of all radioactive peaks.

Undecay-corrected yield is determined as the ratio of the activity of the final liquid composition product measured using an activity meter to ^{the 18} F starting activity.

The method of measuring clarity is as follows: observe the solution to be tested under a clarity detector and compare it with a standard turbidity solution to determine the clarity of the solution.

The method for measuring insoluble particles is as follows: use the photoresist method to measure the insoluble particles content of the solution under an insoluble particle detector, and the unit is particles/mL.

The determination method of radiochemical purity 0h is: using HPLC sample injection analysis, the ratio of the radioactive peak area of the product to the peak area of all radioactive peaks.

The measurement method of 6-hour stability (radiochemical purity index) is: after the final product is placed at room temperature for 6 hours, HPLC is used for sample injection analysis, and the ratio of the radioactive peak area of the product to the peak area of all radioactive peaks.

Table 4 Experimental effect data of Examples 1-18 and Comparative Example 1

Compared with Example 1, Example 2-3 increases the dosage of compound I tert-butyl ester precursor, and its yield and labeling rate increase, but the radiochemical purity does not change much.

Compared with Example 3, in Examples 4 and 5, when the dosage of compound I tert-butyl ester precursor is constant and the ¹⁸ F ion starting activity is increased, the yield and labeling rate change little, and the radiochemical purity changes. Not big.

Compared with Example 5, Examples 6 and 7 changed the volume ratio of trifluoroacetic acid and acetonitrile in the reaction step of removing tert-butyl ester. As the amount of trifluoroacetic acid increases, the yield and labeling rate increase.

Compared with Example 5, Example 8 and Example 9 changed the ratio of mobile phase ethanol and water. The yield and labeling rate of Example 8 were both better than those of Example 5. The yield and labeling rate of Example 9 were The yield and labeling rate are both worse than those of Example 5. When the ethanol content in the mobile phase is greater, the peak time is faster, the preparation time is shorter, and the yield will increase, but radiochemistry Purity will be reduced.

Compared with Example 5, Examples 10-12 changed the content of vitamin C in the mobile phase, increased the yield and labeling rate, and improved the radiochemical purity.

Compared with Example 5, in Examples 13-15, when vitamin C is not included in the mobile phase or vitamin C is replaced with other substances, such as L-glutathione or thiourea, the yield and labeling rate decrease. Radiochemistry Purity is reduced.

In Example 16, in the reaction step of removing tert-butyl ester, a mixed system of trifluoroacetic acid and methylene chloride was used. The yield and labeling rate were low, and the radiochemical purity was not high.

Compared with Example 5, in Examples 17-18, vitamin C was replaced with gentisic acid in the mobile phase, or a mixture of vitamin C and gentisic acid, the yield and labeling rate were equivalent, and the radiochemical purity was equivalent. Gentisic acid It has the same effect as vitamin C in the mobile phase.

In Comparative Example 1, a mixed system of acetonitrile and water was used as the mobile phase, which required an additional C18 column purification step, increased the total preparation time, low yield and labeling rate, and low radiochemical purity.

Table 5 Experimental effect data of Examples 21-27 and Comparative Examples 2-8

In Examples 21-23, the effect of the amount of polysorbate 80 on the stability and radiochemical purity was investigated. As the amount of polysorbate 80 increased, the stability and radiochemical purity of the compound I liquid composition did not change. big.

Compared with Example 21, Examples 24-26 change the dosage of vitamin C. As the dosage of vitamin C increases, the stability and radiochemical purity are improved to a certain extent. When the concentration of vitamin C is between 1mg/mL-10mg/ mL, there is not much change. When the amount of vitamin C increases, the pH value of the solution will decrease, making it too acidic, which is not good for human health.

As a liquid composition containing compound I, that is, a liquid composition of trans-2-(2-(5-(fluoro[ ^18F ])tridecyl)cyclopropyl)acetic acid, the liquid is required to be clear and transparent. In addition, It is required that the number of insoluble particles with a particle size of ≥10 μm should be less than 1,200 particles/mL. Compared with Example 21, in Comparative Examples 2-4, the poly After sorbate 80 is replaced with polyethylene glycol 400 or propylene glycol, or when polysorbate 80 is not used, the clarity (should be clear) and insoluble particles (≥10 μm should be less than 1200 particles/mL) do not meet the requirements.

Compared with Example 21, in Comparative Examples 5-8, when vitamin C is not used or vitamin C is replaced by L-glutathione, thiourea or sodium metabisulfite, the stability and radiochemical purity are reduced.

Compared with Example 21, in Example 27, when vitamin C is replaced by gentisic acid, the stability and radiochemical purity are comparable to Example 21, and the ability of gentisic acid to stabilize Compound I in liquid compositions is comparable. .

On the other hand, for compound I, the radiochemical purity needs to be ≥90%, and <90% is inappropriate and cannot be used.

Although this case has been disclosed through embodiments, they are not intended to limit this case. Anyone with ordinary knowledge in the technical field can make some changes and modifications without departing from the spirit and scope of this case. Therefore, the protection of this case The scope shall be determined by the attached patent application scope.

Claims (14)

A method for preparing a liquid composition of Compound I, wherein: The crude product containing compound I is purified by high performance liquid chromatography; Wherein, the mobile phase used in the high performance liquid chromatography purification step includes ethanol and water; Compound I is trans-2-(2-(5-fluoro[ ^18F ]tridecyl)cyclopropyl)acetic acid. The preparation method according to claim 1, wherein, In the HPLC purification step: In the mobile phase, the ethanol is 2-5 parts by volume relative to 1 part by volume of water. The preparation method according to claim 1, wherein, In the HPLC purification step: The mobile phase also includes vitamin C and/or gentisic acid. The preparation method according to claim 3, wherein, In the HPLC purification step: In the mobile phase, the added amount of vitamin C is 0.1 mg/mL-10 mg/mL. The preparation method according to claim 4, wherein, In the mobile phase, the added amount of vitamin C is 0.1 mg/mL-5 mg/mL. The preparation method according to claim 3, wherein, In the HPLC purification step: In the mobile phase, the added amount of gentisic acid is 0.1 mg/mL-10 mg/mL. The preparation method according to claim 6, wherein, In the mobile phase, the added amount of gentisic acid is 0.1 mg/mL-5 mg/mL. The preparation method according to claim 1, wherein, The chromatographic column is a silica gel column. The preparation method according to claim 1, wherein, Before the high-performance liquid chromatography purification step, a nucleophilic substitution reaction step and a de-tert-butyl esterification reaction step are also included: Nucleophilic substitution reaction: Mix the activated ¹⁸ F ions with a solution containing the precursor of Compound I tert-butyl ester to perform a nucleophilic substitution reaction to obtain an intermediate product solution containing Compound I tert-butyl ester; The precursor of compound I tert-butyl ester is trans-tert-butyl 2-(2-(5-(methanesulfonyloxy)tridecyl)cyclopropyl)acetate; Compound I tert-butyl ester is trans-2-(2-(5-fluoro[ ^18F ]tridecyl)cyclopropyl)acetate tert-butyl ester; De-tert-butyl esterification reaction: Add an acidic solvent to the intermediate product solution of the above compound I tert-butyl ester to perform the de-tert-butyl esterification reaction to obtain a product containing compound I. The preparation method according to claim 9, wherein, In nucleophilic substitution reactions, The starting activity ratio range of compound I tert-butyl ester precursor/ ¹⁸ F is (0.2-5):1; Wherein, the weight unit of the compound I tert-butyl ester precursor is mg, and the unit of initial activity is Ci. The preparation method according to claim 10, wherein, The initial activity of ¹⁸ F is 0.09Ci-11Ci. The preparation method according to claim 9, wherein, In the de-tert-butyl esterification reaction, The acidic solvent is a mixed system of trifluoroacetic acid and acetonitrile. The preparation method according to claim 12, wherein, Relative to 1 part by volume of acetonitrile, trifluoroacetic acid is 0.4-2 parts by volume. Use of the compound I liquid composition prepared according to the method of any one of claims 1 to 13 in preparing a PET imaging agent for myocardial metabolism.