CN211999536U - A [ 2 ]18F]Automatic preparation facilities of D3FSP - Google Patents

Abstract

The utility model relates to a [ alpha ], [ alpha¹⁸F]An automatic preparation device of D3FSP belongs to the technical field of drug synthesis, and comprises a fluorination reaction unit, a fluorination reaction liquid storage unit, a deprotection reaction liquid storage unit and a purification unit; the fluorination reaction unit is respectively connected with the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit through a pipeline and a valve; the fluorination reaction unit is connected with the solid-phase extraction column through a pipeline and a valve; the solid phase extraction column is respectively connected with a waste liquid bottle and a transfer bottle through valves, and the transfer bottle is connected with a product bottle through a valve. The utility model discloses a¹⁸F]An automatic preparation apparatus of D3FSP, which allows the automatic preparation of the radioactive probe [ 2 ]¹⁸F]The D3FSP is successfully realized, the preparation efficiency and the stability are improved, the standardized production is realized, simultaneously, the operator is protected from radiation damage, and the kit is a novel Abeta plaque targeted radioactive probe¹⁸F]The clinical application of the D3FSP lays a foundation.

Description

A [ 2 ]18F]Automatic preparation facilities of D3FSP

Technical Field

The utility model relates to an automatic preparation of positron emission computed tomography (PET) medicine, a purification method of a solid phase extraction column and an automatic synthesis device, in particular to a radioactive probe-, [ alpha ] beta plaque targeting¹⁸F]D3 FSP's automatic preparation facilities belongs to the synthetic technical field of medicine.

Background

Positron Emission Tomography (PET) technology is the most advanced technology of contemporary imaging, which is an imaging technology for non-invasive, dynamic and quantitative evaluation of physiological and biochemical changes in vivo by using positron-emitting drugs as molecular probes. PET has been widely used in differential diagnosis and monitoring of therapeutic effects of tumors, cardiovascular and cerebrovascular diseases, neurological diseases, and the like. Positron medicine is the key to promote the development of PET imaging technology, and is a medicine labeled with positron emitting radionuclide and capable of combining with target specifically after entering human body.

With the social progress and the aging of the population, the central neurodegenerative diseases become important social problems affecting the health level and the quality of life of the population in China. Degenerative diseases of the central nervous system are a general term for a group of diseases caused by chronic progressive degeneration of central nervous tissue, and mainly include Alzheimer Disease (AD), Parkinson Disease (PD), Huntington Disease (HD), and the like. The prevalence of AD is the leading cause of progressive neurodegenerative disease, with clinical characteristic pathological changes including intracellular neurofibrillary tangles (NFTs), extracellular amyloid deposits (SP, commonly known as senile plaques), and the like. The main component of SP amyloid deposit is beta-amyloid (A beta), which is considered to play an important role in the pathogenesis of AD, and at present, the 3 AD diagnostic positron imaging agents approved by the U.S. Food and Drug Administration (FDA) all use A beta as a target.

Spring of 2012, FDA formal approval¹⁸F]AV-45 is marketed in the United states, the first quiltFDA approved positron AD imaging agents are approved by the european medical regulatory agency for the diagnosis and therapeutic monitoring of AD disease the next year. According to literature reports [ Choi S, Golding G, Zhuang Z, Zhang W, Lim N, Hefti F, Benedum T, Kilbourn M, Skovronsky D, Kung H.precrinal properties of 18F-AV-45: a PET agent for A β -plants in the bridge.J.Nucl.Med.2009; 50:1887-94]In vivo injection of the "alpha" [ solution ] in a mouse¹⁸F]After AV-45 and 30min, the mother probe 2¹⁸F]AV-45 accounts for only 30% of the total radioactivity in the serum, and the balance is radioactive metabolite, respectively¹⁸F]The biological half-life of AV-45 is less than 30 min. A study showed that about 50% of the radioactive metabolite in the serum is [ alpha ], [ beta ], [¹⁸F]AV-160, consisting of¹⁸F]N-CH of AV-45₃The site is formed by demethylation, and the specific structure is as follows:

the results of the in vitro binding experiment show that¹⁸F]AV-160 has a low affinity for A β and is about [ alpha ], [ beta ]¹⁸F]1/20 of AV-45, but¹⁸F]AV-160 can cross the Blood Brain Barrier (BBB), bind to non-target areas in the brain, reduce the target to non-target ratio, and thereby reduce the contrast of PET visualizations. For slowing down¹⁸F]The metabolism speed of AV-45 in vivo, Hank F.Kung topic group, N-CD, which is a C-H bond replaced by a C-D bond with strong bond energy₃In place of N-CH₃Synthesizing a novel radioactive probe [ 2 ] targeting an Abeta plaque¹⁸F]D3FSP, structural comparison as follows:

since the physical half-life of the radionuclide F-18 is only 109.8min, the radioactive probe [ 2 ]¹⁸F]D3FSP needs to be prepared on-the-fly and completed as quickly and efficiently as possible.

The literature published in 2019 [ Yao X, ZHa Z, ZHao R, Choi S,

K,Liu F,Zhu L,Kung H.Optimization of solid-phase extraction(SPE)in the preparation of[18F]D3FSP:A new PET imaging agent for mapping Aβplaques.[J].Nuclear Medicine&Biology,2019,71:54-64]a "2" is reported¹⁸F]The preparation method of the D3FSP comprises the following steps: the precursor compound undergoes nucleophilic substitution reaction in the presence of a phase transfer catalyst to obtain a labeled intermediate, undergoes deprotection reaction under acidic conditions, and is purified and formulated to obtain the product¹⁸F]D3FSP product. However, the inventor finds that: when the radioactivity initially charged is increased¹⁸F]The radioactivity of the D3FSP product did not increase because a significant portion of the product was purified during the purification step¹⁸F]The D3FSP product enters the waste liquid bottle along with the leacheate, resulting in¹⁸F]Very low radioactivity in D3FSP products (<1%) the activity of radiopharmaceuticals prepared for clinical imaging generally reaches the curie (Ci) level, and it is clear that the above methods do not meet the requirements of clinical applications.

In order to protect operators from radiation damage and reduce the irradiation dose, the radiopharmaceutical preparation process needs to be completed by an automatic synthesis device. In addition, the automated preparation of the radiopharmaceutical is beneficial to realizing the standardization and repeatability of the preparation process and ensuring the safety and effectiveness of the radiopharmaceutical preparation. Thus, the term¹⁸F]The automated preparation of D3FSP is a prerequisite for its clinical application.

Granted on 19/4/2017, application number 201510654263.9, entitled "A composition¹⁸The Chinese invention patent of F- (2S,4R) -4-fluoro-L-glutamine automatic preparation method and device thereof discloses¹⁸An automatic preparation device of F- (2S,4R) -4-fluoro-L-glutamine. The device has the advantages that the device is provided with two heating reaction systems, can well realize the preparation of the radioactive drug needing two-pot reaction, has the defects of large volume, large space occupation and no contribution to the preparation of the radioactive drug placed in a hot chamber; in addition, the device needs two sets of control systems, is complex to operate and is not easy to carry out 'fool' automatic production.

Zhulin et al published in NorthArticle on the university of Beijing university Natural science bulletin (2011- "¹⁸Development and application of a BNU F-A2 type automatic F positron medicine synthesis device discloses a BNU F-A2 type for a laboratory¹⁸An automatic synthesis device for F positron drugs. The device has the advantages that the clamp type electromagnetic valve is adopted, so that the prepared reagents all flow in the pipeline, and the residues in the valve body and the corrosion to the valve body are avoided; but has the disadvantages that the environment is not friendly, and the volatile organic reagent is directly discharged into the air in the preparation process; in addition, the device is not flexible enough to perform only specific steps and the number of radiopharmaceuticals that can be prepared is limited.

Thus, a highly efficient, stable, safe, environment-friendly [ alpha ], [ beta ] -an¹⁸F]The D3FSP automatic preparation method and the device thereof become a technical problem which needs to be solved urgently in the field.

SUMMERY OF THE UTILITY MODEL

One of the purposes of the utility model is to provide a high-efficiency, stable, safe and environment-friendly¹⁸F]An automated preparation method of D3 FSP.

The above object of the utility model is achieved through the following technical scheme:

preparation by using automatic device¹⁸F]A method of D3FSP, comprising the steps of:

(1) fluorination reagents¹⁸F^-Trapping of ions

Make it contain¹⁸F^-Passing the aqueous solution of ions through an anion exchange column¹⁸F^-Adsorbing the ions on an anion exchange column;

(2) the nucleophilic reaction active reagent K222/K [ solution ]¹⁸F]Preparation of

Adsorbing step (1) on an anion exchange column with a solution containing a phase transfer catalyst¹⁸F^-The ion elution enters a fluorination reaction unit, and the solvent in the fluorination reaction unit is evaporated under the conditions of inert gas flow purging and heating condition to obtain the dry nucleophilic reaction active reagent K222/K [ ]¹⁸F]；

(3) Fluorination reaction

Dissolving the precursor compoundDissolved in an aprotic solvent, and added to the dried nucleophilic reaction active reagent K222/K [ prepared in step (2)¹⁸F]In the method, a fluorination reaction unit is sealed, and after reaction for a certain time under a heating condition, the reaction mixture solution containing an intermediate is obtained by cooling;

(4) deprotection reaction of intermediate

Adding sulfuric acid with a certain concentration into the reaction mixed solution containing the intermediate obtained in the step (3), sealing the heating reaction unit, carrying out heating reaction for a certain time, and cooling to obtain a mixed solution of a crude product;

(5) product purification by solid phase extraction column

Adding a sodium hydroxide (NaOH) solution into the mixed solution of the crude product obtained in the step (4), neutralizing the mixed solution, enabling the mixed solution to pass through a solid-phase extraction column, washing the solid-phase extraction column by using water, washing the solid-phase extraction column for the second time by using acetonitrile/water solution with a certain ratio after washing, and washing the solid-phase extraction column for the third time by using water after washing to obtain a purified solid-phase extraction column;

(6) product preparation

Using ethanol to dissolve the [ alpha ]¹⁸F]Eluting the D3FSP product from the purified solid phase extraction column in the step (5) into physiological saline, and passing through a sterile filter membrane to obtain the preparation [ 2 ]¹⁸F]D3FSP。

The preparation reaction equation is as follows:

preferably, in step (1), the anion exchange column is a QMA column, and further preferably, the QMA column is a 4- (4-methylpiperidine) pyridine anion exchange resin column.

Preferably, in the step (2), the phase transfer catalyst solution is K222/K₂CO₃And (3) a solution containing a proper amount of acetonitrile and water.

Preferably, in the step (2), the blow-drying is performed under the conditions of anhydrous acetonitrile and 110 ℃, and the inert gas is nitrogen or argon.

Preferably, in step (3), the heating temperature is 120 ℃, the reaction time is 10min, the aprotic solvent is DMSO, and the precursor compound is (E)4- (2- (6- (2- (2- (2-fluoroethoxy) ethoxy) pyridin-3-yl) vinyl) -N- (methyl-D3) aniline.

Preferably, in the step (4), the concentration of the sulfuric acid is 2mol/L, the reaction temperature is 100 ℃, and the reaction time is 5 min.

Preferably, in the step (5), the concentration of the sodium hydroxide (NaOH) solution is 0.4mol/L, the solid phase extraction column is an Oasis HLB column, the volume of the water for the first washing is 10mL, the acetonitrile/water ratio is 35%, and the volume of the water for the third washing is 20 mL.

Preferably, in step (6), the volume of ethanol is 2mL, and the volume of physiological saline is 18 mL.

Another object of the present invention is to provide the above [ 2 ]¹⁸F]D3 FSP's automatic preparation facilities.

The above object of the utility model is achieved through the following technical scheme:

a [ 2 ]¹⁸F]D3FSP automated preparation device, its characterized in that: the device comprises a fluorination reaction unit, a fluorination reaction liquid storage unit, a deprotection reaction liquid storage unit and a purification unit; the fluorination reaction unit is respectively connected with the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit through a pipeline and a valve; the fluorination reaction unit is also connected with an oxygen 18 water recovery bottle and an anion exchange column through a pipeline and a valve; the anion exchange column is loaded with K222/K through a pipeline and a valve₂CO₃The penicillin bottles of the solution are connected; the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit are connected with each other through pipelines and valves; the fluorination reaction unit is connected with the low-temperature negative-pressure waste gas collecting unit through a pipeline and a valve; the fluorination reaction unit is connected with inert gas (such as N) through a gas pipeline and a valve₂) A supply device connection; the fluorination reaction unit is connected with the solid-phase extraction column through a pipeline and a valve; the solid phase extraction column is respectively connected with a waste liquid bottle and a transfer bottle through valves, and the transfer bottle is connected with a product bottle through a valve。

Preferably, the fluorination reaction liquid storage unit comprises a K222/K loading unit₂CO₃A penicillin bottle 1 for the solution, a penicillin bottle 2 loaded with anhydrous acetonitrile, a penicillin bottle 3 loaded with anhydrous acetonitrile and a penicillin bottle 4 loaded with a reaction precursor compound DMSO solution; one end of the reaction vessel is connected with the reaction vessel 14 through a fourth solenoid valve 23, a fifth solenoid valve 24 and a sixth solenoid valve 25, and the other end is connected with an inert gas (such as N) through a third solenoid valve 22 and a gas pipeline₂) The supply device is connected.

Preferably, the deprotection reaction reservoir unit comprises a load H₂SO₄One end of each of the penicillin bottle 5 for solution, the penicillin bottle 6 for NaOH solution loading and the penicillin bottle 7 for sterile injection water loading is connected with the reaction bottle 14 through an eighth electromagnetic valve 27, a ninth electromagnetic valve 28 and a tenth electromagnetic valve 29, and the other end is connected with inert gas (such as N) through a seventh electromagnetic valve 26 and a gas pipeline₂) The supply device is connected.

Preferably, the purification unit comprises a penicillin bottle 8 loaded with an acetonitrile/water mixed solution, a penicillin bottle 9 loaded with sterilized water for injection and a penicillin bottle 10 loaded with medicinal grade ethanol; one end of the reaction vessel is connected with the reaction vessel 14 through a fourteenth solenoid valve 33, a fifteenth solenoid valve 34, a sixteenth solenoid valve 35 and a twelfth solenoid valve 31, and the other end is connected with an inert gas (such as N) through a thirteenth solenoid valve 32 and a gas pipeline₂) The supply device is connected.

Preferably, the transfer bottle is a penicillin bottle 16 loaded with sterilized normal saline for injection, and the product bottle is a sealed penicillin bottle 17 loaded with a sterile filter membrane 43.

Preferably, the fluorination reaction unit (reaction flask 14) comprises a temperature-controllable resistance wire 41 and a small electric fan 42 providing ventilation air.

Preferably, the low-temperature negative-pressure waste gas collecting unit comprises a cold trap 13, a dewar 40 loaded with liquid nitrogen and a vacuum pump 12 for providing negative pressure, wherein the vacuum pump 12 is connected with the cold trap 13 through a twenty-third solenoid valve 39, and the cold trap 13 is connected with the reaction flask 14 through a nineteenth solenoid valve 38.

Preferably, the anion exchange column is a QMA cartridge 18 and the solid phase extraction column is an Oasis HLB cartridge 19.

Has the advantages that:

the utility model discloses a¹⁸F]An automatic preparation apparatus of D3FSP, which allows the automatic preparation of the radioactive probe [ 2 ]¹⁸F]The D3FSP is successfully realized, the preparation efficiency and the stability are improved, the standardized production is realized, simultaneously, the operator is protected from radiation damage, and the kit is a novel Abeta plaque targeted radioactive probe¹⁸F]The clinical application of the D3FSP lays a foundation.

The invention is further illustrated by the accompanying drawings and the detailed description, which are not meant to limit the scope of the invention.

Drawings

Figure 1 is the utility model¹⁸F]The structure schematic diagram of the D3FSP automatic preparation device.

FIG. 2-1 shows a sample of the present invention prepared in example 1 of the present invention¹⁸F]HPLC result chart of D3FSP product solution.

FIG. 2-2 shows a solution of the polypeptide prepared in example 2 of the present invention¹⁸F]HPLC result chart of D3FSP product solution.

FIG. 3 is a control compound [ 2 ] containing a stabilizing nuclide¹⁹F]HPLC result of D3FSP solution.

1 load K222/K₂CO₃Penicillin bottle for solution

2 penicillin bottle loaded with anhydrous acetonitrile

3 penicillin bottle loaded with anhydrous acetonitrile

4 penicillin bottle for loading reaction precursor compound DMSO solution

5 Loading H₂SO₄Penicillin bottle for solution

6 penicillin bottle loaded with NaOH solution

7 penicillin bottle loaded with sterilized water for injection

8 XiLin bottle of load acetonitrile/water mixed solution

9 penicillin bottle loaded with sterilized water for injection

10 penicillin bottle loaded with medicinal grade ethanol

Vacuum pump 12 of 11 oxygen 18 water recovery bottle

13 cold trap 14 reaction bottle

15 waste liquid bottle 16 transfer bottle

17 product bottle 18 QMA small column

19 Oasis HLB column 20 first three-way solenoid valve

21 second three-way solenoid valve 22 third solenoid valve

23 fourth solenoid valve 24 fifth solenoid valve

25 sixth solenoid valve 26 seventh solenoid valve

27 eighth solenoid valve 28 ninth solenoid valve

29 tenth solenoid valve 30 eleventh solenoid valve

31 twelfth solenoid valve 32 thirteenth solenoid valve

33 fourteenth solenoid valve 34 fifteenth solenoid valve

35 sixteenth solenoid valve 36 seventeenth solenoid valve

37 eighteenth three-way solenoid valve 38 nineteenth solenoid valve

39 twentieth three-way electromagnetic valve 40 Dewar flask

41 resistance wire 42 electric fan

43 sterile filtration Membrane

Detailed Description

In the field of radiopharmaceutical chemistry, for fluorine compounds, the control compound (standard compound) is generally referred to as nonradioactive¹⁹F]The compound of (1), the product (radioactive probe) generally means radioactive [ 2 ]¹⁸F]The compound of (1).

Reagents and consumables:

containing H¹⁸The aqueous solution of F was prepared from sumitomo HM20 medical cyclotron; k222, K₂CO₃Anhydrous acetonitrile, anhydrous DMSO, ethanol, pharmaceutical grade ethanol, physiological saline, and sterile water for injectionigma-Aldrich; anion exchange columns (QMA cartridges) and solid phase extraction columns (Oasis HLB cartridges) were purchased from Waters; sterile filters were purchased from Merck-Millipore.

The precursor compound is provided by professor Hank F.Kung, university of Pennsylvania, laboratory, and has a chemical purity of more than 95%, and is named as (E)4- (2- (6- (2- (2- (2-fluoroethoxy) ethoxy) pyridin-3-yl) vinyl) -N- (methyl-D3) aniline, and the structural formula is shown as follows:

as shown in figure 1, is the utility model¹⁸F]The structure schematic diagram of the D3FSP automatic preparation device, wherein 1 is a penicillin bottle loaded with a K222/K2CO3 solution, 2 is a penicillin bottle loaded with anhydrous acetonitrile, 3 is a penicillin bottle loaded with anhydrous acetonitrile, 4 is a penicillin bottle loaded with a reaction precursor compound DMSO solution, and 5 is a penicillin bottle loaded with H₂SO₄A penicillin bottle of the solution, 6 is a penicillin bottle loaded with NaOH solution, 7 is a penicillin bottle loaded with sterilized water for injection, 8 is a penicillin bottle loaded with acetonitrile/water mixed solution, 9 is a penicillin bottle loaded with sterilized water for injection, 10 is a penicillin bottle loaded with medicinal grade ethanol, 11 is an oxygen 18 water recovery bottle, 12 is a vacuum pump, 13 is a cold trap, 14 is a reaction bottle, 15 is a waste liquid bottle, 16 is a transfer bottle, 17 is a product bottle, 18 is a QMA small column, 19 is an Oasis HLB small column, 20 is a first three-way solenoid valve, 21 is a second three-way solenoid valve, 22 is a third solenoid valve, 23 is a fourth solenoid valve, 24 is a fifth solenoid valve, 25 is a sixth solenoid valve, 26 is a seventh solenoid valve, 27 is an eighth solenoid valve, 28 is a ninth solenoid valve, 29 is a tenth solenoid valve, 30 is an eleventh solenoid valve, 31 is a twelfth solenoid valve, 32 is a thirteenth solenoid valve, 33 is a fourteenth solenoid valve, 34 is a fifteenth electromagnetic valve, 35 is a sixteenth electromagnetic valve, 36 is a seventeenth electromagnetic valve, 37 is an eighteenth electromagnetic valve, 38 is a nineteenth electromagnetic valve, 39 is a twentieth electromagnetic valve, 40 is a dewar flask, 41 is a resistance wire, 42 is an electric fan, and 43 is a sterile filter membrane.

The term "2" of example 1¹⁸F]The D3FSP automatic preparation device comprisesThe device comprises a fluorination reaction unit, a fluorination reaction liquid storage unit, a deprotection reaction liquid storage unit and a purification unit; the fluorination reaction unit is respectively connected with the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit through a pipeline and a valve; the fluorination reaction unit is also connected with an oxygen 18 water recovery bottle 11 and an anion exchange column QMA column 18 through pipelines and valves; the anion exchange column QMA column 18 is loaded with K222/K through a pipeline and a valve₂CO₃Connecting penicillin bottles 1 of the solution; the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit are connected with each other through a pipeline and a valve; the fluorination reaction unit is connected with the low-temperature negative-pressure waste gas collecting unit through a pipeline and a valve; the fluorination reaction unit is connected with inert gas (such as N) through a gas pipeline and a valve₂) A supply device connection; the fluorination reaction unit is connected with a solid phase extraction column Oasis HLB small column 19 through a pipeline and a valve; the solid phase extraction column Oasis HLB small column 19 is respectively connected with the waste liquid bottle 15 and the transfer bottle 16 through valves, and the transfer bottle 16 is connected with the product bottle 17 through valves.

The method comprises the following specific steps: the fluorination reaction liquid storage unit comprises a K222/K loading unit₂CO₃A penicillin bottle 1 for the solution, a penicillin bottle 2 loaded with anhydrous acetonitrile, a penicillin bottle 3 loaded with anhydrous acetonitrile and a penicillin bottle 4 loaded with a reaction precursor compound DMSO solution; one end of each bottle is respectively connected with the reaction bottle 14 through a fourth electromagnetic valve 23, a fifth electromagnetic valve 24, a sixth electromagnetic valve 25 and a liquid pipeline, and the other end is connected with inert gas (such as N) through a third electromagnetic valve 22 and a gas pipeline₂) A supply device connection;

the deprotection reaction liquid storage unit comprises a load H₂SO₄A penicillin bottle 5 for solution, a penicillin bottle 6 for loading NaOH solution and a penicillin bottle 7 for loading sterilized water for injection, wherein one end of each bottle is respectively connected with the reaction bottle 14 through an eighth electromagnetic valve 27, a ninth electromagnetic valve 28, a tenth electromagnetic valve 29 and a liquid pipeline, and the other end is connected with inert gas (such as N) through a seventh electromagnetic valve 26 and a gas pipeline₂) A supply device connection;

the purification unit comprises a penicillin bottle 8 loaded with acetonitrile/water mixed solution, a penicillin bottle 9 loaded with sterilized water for injection and a penicillin bottle loaded with medicinal ethanol10; one end of each bottle is respectively connected with the reaction bottle 14 through a fourteenth electromagnetic valve 33, a fifteenth electromagnetic valve 34, a sixteenth electromagnetic valve 35, a twelfth electromagnetic valve 31 and a liquid pipeline and through the twelfth electromagnetic valve 31; the other end is connected with inert gas (such as N) through a thirteenth electromagnetic valve 32 and a gas pipeline₂) A supply device connection;

the transfer bottle 16 is a penicillin bottle loaded with sterilized injection physiological saline, and the product bottle 17 is a sealed penicillin bottle loaded with a sterile filter membrane;

the reaction flask 14 comprises a temperature-controllable resistance wire 41 and an electric fan 42 for providing ventilation air;

the low-temperature negative-pressure waste gas collecting unit comprises a cold trap 13, a Dewar flask 40 loaded with liquid nitrogen and a vacuum pump 12 for providing negative pressure, the vacuum pump 12 is connected with the cold trap 13 through a twenty-third electromagnetic valve 39, and the cold trap 13 is connected with the reaction flask 14 through a nineteenth electromagnetic valve 38;

the anion exchange column was a QMA cartridge 18 and the solid phase extraction column was an Oasis HLB cartridge 19(3 cc).

Example 1

[¹⁸F]Preparation of D3FSP (solid phase extraction column purification)

The method comprises the following specific steps:

1) preparation of H-containing from Nippon sumitomo HM20 medical cyclotron¹⁸An aqueous solution of F, which was passed through a QMA cartridge in one portion, and then 1.1mL of an eluent (11mg K222 and 2mg K) was used₂CO₃Dissolved in 0.93mL of acetonitrile and 0.17mL of water) of¹⁸F]After the ions were eluted, the resulting solution was purged with nitrogen (N) at 110 deg.C₂) Blow-drying, and sequentially adding 1mL of anhydrous acetonitrile (2 times in total) to continue blow-drying to obtain the dried [ 2 ]¹⁸F]KF/K222；

2) The precursor compound (E), 4- (2- (6- (2- (2- (2-fluoroethoxy) ethoxy) pyridin-3-yl) vinyl) -N- (methyl-D3) aniline, was dissolved in 1mL of anhydrous DMSO, and the dried product prepared in step 1) (2)¹⁸F]In KF/K222, after fully shaking and shaking up, placing at 120 ℃, and sealing for fluorination reaction for 10 minutes;

3) the reaction mixture was cooled for 1 minute, and 1mL of 2mol/L H was added₂SO₄Solution, deprotection reaction is carried out for 5 minutes at the temperature of 100 ℃, then 10mL of NaOH solution with the concentration of 0.4mol/L is added to neutralize the reaction solution, and the mixed solution passes through an Oasis HLB column;

4) flushing the Oasis HLB column sequentially with 10mL water for injection, 6mL of 35% acetonitrile/water solution (vol/vol) and 20mL water for injection;

5) 2mL of pharmaceutical grade ethanol is used for dissolving¹⁸F]Eluting the D3FSP product from the Oasis HLB column into 18mL of physiological saline, and passing the mixed solution through a sterile filter membrane to obtain the product of [ 2 ], [ 2 ]¹⁸F]D3FSP formulation.

The preparation result is as follows: the total preparation time is about 45 minutes, the final preparation yield is 27%, the activity of the preparation is 1.5-3mCi, the radiochemical purity is more than 95%, and the chemical impurity content in the preparation is less than 50 mu g.

Prepared [ alpha ], [ alpha ]¹⁸F]D3FSP structure validation:

the product obtained in example 1 [ alpha ], [ alpha ]¹⁸F]D3FSP and a control Compound¹⁹F]D3FSP, co-injected under the same conditions and analyzed by HPLC, as shown in FIG. 2-1, is the product prepared in example 1 of the present invention¹⁸F]HPLC profile of D3 FSP; as shown in FIG. 3, is a control compound of the present invention¹⁹F]HPLC profile of D3 FSP.

Analytical HPLC conditions were as follows:

a chromatographic column: luna C₁₈，150x 4.6mm，5μm；

Chromatograph: agilent 1260 (equipped with uv detector and radioactivity detector);

mobile phase: a: 10mM ammonium formate solution B: acetonitrile, flow rate: 1 mL/min;

ultraviolet detection wavelength: 350nm, run time: and 20 min.

Gradient conditions are given in Table 1 below

Table 1: gradient condition

The retention time is confirmed by comparing the retention time, so that the retention time of the two is consistent, and the method canTo prepare the [ alpha ], [ alpha ] is prepared successfully¹⁸F]The D3FSP product (see fig. 2-1 and 3).

Example 2

[¹⁸F]Automated preparation of D3FSP

The term "in example 1¹⁸F]The preparation parameters of the D3FSP are transmitted to the automatic synthesizer in the utility model through programming, and the automatic synthesizer is utilized to carry out¹⁸F]Automated preparation of D3 FSP.

a)[¹⁸F]Preparation work for automated preparation of D3FSP

At load K222/K₂CO₃Solution penicillin bottle 1 is loaded with K222/K₂CO₃A solution; respectively loading 1mL of anhydrous acetonitrile in a penicillin bottle 2 loaded with the anhydrous acetonitrile and a penicillin bottle 3 loaded with the anhydrous acetonitrile; 1mL of anhydrous DMSO solution containing 1mg of precursor compound is loaded in a penicillin bottle 4 loaded with a reaction precursor compound DMSO solution; in loading H₂SO₄1mL of H with the concentration of 2mol/L is loaded in a solution penicillin bottle 5₂SO₄A solution; loading 10mL of NaOH solution with the concentration of 0.4mol/L into a penicillin bottle 6 loaded with the NaOH solution; loading 10mL of sterile water for injection into a penicillin bottle 7 loaded with water for injection; 6mL of acetonitrile/water solution (volume ratio) with the concentration of 35 percent is loaded in a penicillin bottle 8 loaded with the acetonitrile/water solution; 20mL of sterilized water for injection is loaded in a vial 9 for loading water for injection; loading 2mL of medicinal ethanol into a penicillin bottle 10 loaded with the medicinal ethanol; loading 18mL of physiological saline into the transfer bottle 16;

the normally open port of the first three-way electromagnetic valve 1 is connected with the Sumitomo HM20 medical cyclotron¹⁸F]The ionic solution delivery pipe is connected; loading the normally closed port of the first three-way electromagnetic valve 1 with K222/K₂CO₃The penicillin bottle 1 of the solution is connected, and a common port of the first three-way electromagnetic valve 1 is connected with the QMA small column 18; connecting a normally open port of the second three-way electromagnetic valve 2 with the oxygen 18 water recovery bottle 11; a normally closed port of the second three-way electromagnetic valve 2 is connected with the reaction bottle 14, and a common port of the second three-way electromagnetic valve 2 is connected with the QMA small column 18;

a normally open port of the seventeenth three-way electromagnetic valve 36 is connected with the waste liquid bottle 15, a common port of the seventeenth three-way electromagnetic valve 36 is connected with the Oasis HLB column 19, and a normally closed port of the seventeenth three-way electromagnetic valve 36 is connected with the transfer bottle 16;

connecting the third solenoid valve 22, the seventh solenoid valve 26 and the thirteenth solenoid valve 32 with the inert gas supply apparatus;

the Oasis small column 19 is respectively connected with the fourteenth electromagnetic valve 33, the fifteenth electromagnetic valve 34 and the sixteenth electromagnetic valve 35;

loading an anion exchange column, namely a QMA small column 18 between a first three-way electromagnetic valve 20 and a second three-way electromagnetic valve 21; a solid phase extraction column, namely an Oasis HLB column 19, is loaded between the twelfth electromagnetic valve 31 and the seventeenth electromagnetic valve 36; loading a sterile filter membrane 43 at the front end of the product bottle;

b)[¹⁸F]automated preparation of D3FSP

The synthesis device is started to complete the preparation work, and the process is carried out¹⁸F]The automatic preparation of the D3FSP comprises the following specific steps:

1) preparation of H-containing from Nippon sumitomo HM20 medical cyclotron¹⁸F, which is caused to pass through the normally open port of the first three-way solenoid valve 20, the QMA small column 18 and the normally open port of the second three-way solenoid valve 21 in this order¹⁸F]The ions are adsorbed on the QMA small column 18, and the leached waste liquid enters an oxygen 18 water recovery bottle 11;

2) opening the twenty-third electromagnetic valve 39 and the nineteenth electromagnetic valve 38 and the vacuum pump 12 to provide negative pressure for the reaction bottle 14, then opening the normally closed ports of the first three-way electromagnetic valve 20 and the second three-way electromagnetic valve 21, and loading K222/K₂CO₃K222/K in penicillin bottle 1 of solution₂CO₃Solution of alpha-cyclodextrin¹⁸F]Leaching ions from the QMA small column 18 into the reaction flask 14, heating the reaction flask 14 to 110 ℃, evaporating the leacheate, and then closing the normally closed ports of the first three-way electromagnetic valve 20 and the second three-way electromagnetic valve 21;

3) the third electromagnetic valve 22 and the fourth electromagnetic valve 23 are opened, and the anhydrous acetonitrile in the penicillin bottle 2 loaded with the anhydrous acetonitrile is added to the solution containing the evaporated water¹⁸F]The KF/K222 residue in the reaction flask 14, at 110 degrees C to dry again, then closed the fourth solenoid valve 23;

4) opening the fifth electromagnetic valve 24, adding anhydrous acetonitrile in the penicillin bottle 3 loaded with the anhydrous acetonitrile into the reaction bottle 14, evaporating to dryness again at 110 ℃ (evaporating to dryness for the third time), and then closing the fifth electromagnetic valve 24;

5) opening a sixth electromagnetic valve 25, adding the anhydrous DMSO solution containing the precursor compound in the penicillin bottle 4 loaded with the anhydrous DMSO solution containing the precursor compound into the reaction bottle 14, then closing the third electromagnetic valve 22, the sixth electromagnetic valve 25, the nineteenth electromagnetic valve 38, the twenty-third electromagnetic valve 39 and the vacuum pump 12 in sequence, sealing the reaction bottle 14, and sealing the fluorination reaction bottle for 10 minutes at 120 ℃;

6) after the reaction is finished, the heating is stopped, the reaction product is cooled for 1 minute, the nineteenth electromagnetic valve 38 and the twenty-third electromagnetic valve 39 are opened, the seventh electromagnetic valve 26 and the eighth electromagnetic valve 27 are opened, and the reaction product is loaded with H₂SO₄1mL of H with the concentration of 2mol/L in a penicillin bottle 5 of the solution₂SO₄Adding the solution into the reaction bottle 14, then closing the seventh electromagnetic valve 26, the eighth electromagnetic valve 27, the nineteenth electromagnetic valve 38 and the twenty-third electromagnetic valve 39, sealing the reaction bottle 14, and carrying out deprotection reaction for 5 minutes at 100 ℃;

7) after the deprotection reaction is finished, stopping heating, cooling for 1 minute, opening a seventh electromagnetic valve 26, a ninth electromagnetic valve 28, a nineteenth electromagnetic valve 38 and a twentieth electromagnetic valve 39, adding 10mL of NaOH solution with the concentration of 0.4mol/L in the penicillin bottle 6 loaded with the NaOH solution into the reaction bottle 14, neutralizing the reaction solution, then closing the nineteenth electromagnetic valve 38 and the twentieth electromagnetic valve 39, opening a twelfth electromagnetic valve 31, allowing the mixed solution in the reaction bottle 14 to enter the waste liquid bottle 15 through the Oasis HLB column 19 and the seventeenth electromagnetic valve 36, and then closing the ninth electromagnetic valve 28;

8) opening the tenth electromagnetic valve 29 to enable 10mL of sterile water for injection in the penicillin bottle 7 loaded with the sterile water for injection to sequentially flush the reaction bottle 14 and the Oasis HLB column 19, then entering the waste liquid bottle 15, and closing the seventh electromagnetic valve 26, the tenth electromagnetic valve 29 and the twelfth electromagnetic valve 31;

9) opening a thirteenth electromagnetic valve 32 and a fourteenth electromagnetic valve 33, allowing 6mL of 35% acetonitrile/water solution in the penicillin bottle 8 loaded with the acetonitrile/water solution to pass through the Oasis HLB column 19 and enter the waste liquid bottle 15, and then closing the fourteenth electromagnetic valve 33;

10) opening a fifteenth electromagnetic valve 34, enabling 20mL of sterile water for injection in the penicillin bottle 9 loaded with the sterile water for injection to enter the waste liquid bottle 15 through the Oasis HLB small column 19, and then closing the fifteenth electromagnetic valve 34;

11) opening the normally closed ends of the sixteenth electromagnetic valve 35 and the seventeenth three-way electromagnetic valve 36, and allowing 2mL of pharmaceutical grade ethanol in the penicillin bottle 10 loaded with the pharmaceutical grade ethanol to enter the Oasis HLB small column 19¹⁸F]D3FSP is eluted from Oasis HLB column 19 into transfer vial 16;

12) opening the normally closed end of the eighteenth three-way electromagnetic valve 37, the [ 2 ] in the bottle 16 is rotated¹⁸F]The D3FSP product is transferred through a sterile filter membrane 43 into a product bottle 17 to obtain the [ alpha ], [ beta ], [¹⁸F]D3FSP formulation.

13) All solenoid valves are closed.

The preparation result is as follows: the total preparation time is about 40 minutes, the final preparation yield is 30 percent, the activity of the preparation is 300-450mCi, the activity of the preparation is greatly improved, the radiochemical purity is more than 95 percent, and the content of chemical impurities in the preparation is less than 50 mu g.

Prepared [ alpha ], [ alpha ]¹⁸F]D3FSP structure validation:

the product obtained in example 2¹⁸F]D3FSP and a control Compound¹⁹F]D3FSP, co-injected under the same conditions and analyzed by HPLC, as shown in FIG. 2-2, is the product prepared in example 2 of the present invention¹⁸F]HPLC profile of D3 FSP; as shown in FIG. 3, is a control compound of the present invention¹⁹F]HPLC profile of D3 FSP.

Analytical HPLC conditions were as follows:

a chromatographic column: luna C₁₈，150x4.6mm，5μm；

Chromatograph: agilent 1260 (equipped with uv detector and radioactivity detector);

mobile phase: a: 10mM ammonium formate solution B: acetonitrile, flow rate: 1 mL/min;

ultraviolet detection wavelength: 350nm, run time: and 20 min.

Gradient conditions are shown in table 2:

TABLE 2

The retention time is confirmed by comparing the retention times, so that the product can be successfully prepared by using an automatic synthesis apparatus¹⁸F]The D3FSP product (see fig. 2-2 and 3).

The utility model discloses a¹⁸F]The D3FSP automatic preparation device has the advantages that: firstly, a pipe clamping type electromagnetic valve is adopted to control liquid to flow, so that the liquid does not directly contact with a valve body of the electromagnetic valve, and the corrosion of a reagent to the electromagnetic valve and the residue in the valve body are avoided; secondly, a negative pressure system is arranged, so that the evaporation and dehydration speed can be increased, and the preparation time can be shortened; thirdly, the traditional semi-preparative high-pressure liquid chromatography purification method is replaced by the solid-phase extraction small column purification method, which is beneficial to simplifying the preparation process flow and improving the preparation efficiency; fourth, the activity of the formulation is greatly increased.

The utility model discloses a¹⁸F]An automatic preparation apparatus of D3FSP, which allows the automatic preparation of the radioactive probe [ 2 ]¹⁸F]The D3FSP is successfully realized, the preparation efficiency and the stability are improved, the standardized production is realized, simultaneously, the operator is protected from radiation damage, and the kit is a novel Abeta plaque targeted radioactive probe¹⁸F]The popularization and clinical application of the D3FSP lay a foundation.

Claims (10)

1. A [ 2 ]¹⁸F]D3FSP automated preparation device, its characterized in that: the device comprises a fluorination reaction unit, a fluorination reaction liquid storage unit, a deprotection reaction liquid storage unit and a purification unit; the fluorination reaction unit is respectively connected with the fluorination reaction liquid storage unit, the deprotection reaction liquid storage unit and the purification unit through a pipeline and a valve; the fluorination reaction unit is also connected with an oxygen 18 water recovery bottle and an anion exchange column through a pipeline and a valve; the anion exchange column is loaded with K222/K through a pipeline and a valve₂CO₃The penicillin bottles of the solution are connected; the fluorination reaction liquid storage unit and the reactorThe protective reaction liquid storage unit and the purification unit are connected with each other through a pipeline and a valve; the fluorination reaction unit is connected with the low-temperature negative-pressure waste gas collecting unit through a pipeline and a valve; the fluorination reaction unit is connected with an inert gas supply device through a gas pipeline and a valve; the fluorination reaction unit is connected with the solid-phase extraction column through a pipeline and a valve; the solid phase extraction column is respectively connected with the waste liquid bottle and the transfer bottle through valves, and the transfer bottle is connected with the product bottle through valves.

2. The [ 2 ] of claim 1¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the fluorination reaction liquid storage unit also comprises a penicillin bottle loaded with anhydrous acetonitrile, a penicillin bottle loaded with anhydrous acetonitrile and a penicillin bottle loaded with a reaction precursor compound DMSO solution; one end of the reaction bottle is connected with the reaction bottle through a fourth electromagnetic valve, a fifth electromagnetic valve and a sixth electromagnetic valve, and the other end of the reaction bottle is connected with inert gas supply equipment through a third electromagnetic valve and a gas pipeline.

3. The [ 2 ] of claim 2¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the deprotection reaction liquid storage unit comprises a load H₂SO₄One end of each of the penicillin bottle for the solution, the penicillin bottle for loading the NaOH solution and the penicillin bottle for loading the sterilized injection water is connected with the reaction bottle through an eighth electromagnetic valve, a ninth electromagnetic valve and a tenth electromagnetic valve, and the other end of each of the penicillin bottles for the solution, the penicillin bottle for loading the NaOH solution and the penicillin bottle for loading the sterilized injection water is connected with the inert gas supply equipment through a seventh electromagnetic valve and a gas pipeline.

4. The [ 2 ] of claim 3¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the purification unit comprises a penicillin bottle loaded with acetonitrile/water mixed solution, a penicillin bottle loaded with sterilized water for injection and a penicillin bottle loaded with medicinal ethanol; one end of the reaction bottle is connected with the reaction bottle through a fourteenth electromagnetic valve, a fifteenth electromagnetic valve, a sixteenth electromagnetic valve and a twelfth electromagnetic valve, and the other end of the reaction bottle is connected with inert gas supply equipment through a thirteenth electromagnetic valve and a gas pipeline.

5. The [ 2 ] of claim 4¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the transfer bottle is a penicillin bottle loaded with sterilized injection physiological saline.

6. The [ 2 ] of claim 5¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the product bottle is a sealed penicillin bottle loaded with a sterile filter membrane.

7. The [ 2 ] of claim 6¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the anion exchange column is a QMA small column.

8. The [ 2 ] of claim 7¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the solid phase extraction column is an Oasis HLB column.

9. The [ 2 ] of claim 8¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the fluorination reaction unit comprises a temperature-controllable resistance wire and a small electric fan for providing ventilation air.

10. The [ 2 ] of claim 9¹⁸F]D3 FSP's automatic preparation facilities, its characterized in that: the low-temperature negative-pressure waste gas collecting unit comprises a cold trap, a liquid nitrogen loading Dewar flask and a vacuum pump for providing negative pressure, the vacuum pump is connected with the cold trap through a twenty-third electromagnetic valve, and the cold trap is connected with the reaction flask through a nineteenth electromagnetic valve.

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