阅读说明：本技术 N-(2-氟[18f]乙基)多巴胺显像剂及其制备方法 (N- (2-fluoro [18F ] ethyl) dopamine developer and preparation method thereof ) 是由 何玉林 王相成 周伟娜 张国建 白侠 王雪梅 杨宏娟 于 2020-05-05 设计创作，主要内容包括：本发明涉及N-(2-氟[～(18)F]乙基)多巴胺显像剂及其制备方法,该制备方法包括：(1)将回旋加速器生产的氟离子(～(18)F～(-))传入化学合成器捕获在阴离子捕获,洗脱,加乙腈共沸去水；(2)加入式(Ⅱ)的乙腈溶液反应；(3)加入多巴胺的二甲亚砜溶液反应；(4)高效液相分离纯化,收集所需放射性组分N-(2-氟[～(18)F]乙基)多巴胺。本发明所述显像剂与多巴胺转运体和去甲肾上腺素转运体的亲和力强,其能够早期发现心脏交感神经的变化,为心脏疾病的早期诊断及治疗提供有力的证据；本制备方法重复性好,合成产率稳定。(The present invention relates to N- (2-fluoro-) 18 F]Ethyl) dopamine developer and its preparation method, said preparation method comprises: (1) fluorine ions produced by a cyclotron (A) 18 F ‑ ) Transferring into a chemical synthesizer for capturing in anion, eluting, and adding acetonitrile for azeotropic dehydration; (2) adding acetonitrile solution of formula (II) for reaction; (3) adding a dimethyl sulfoxide solution of dopamine for reaction; (4) separating and purifying with high performance liquid phase, and collecting the required radioactive component N- (2-fluoro-) 18 F]Ethyl) dopamine. The imaging agent has strong affinity with a dopamine transporter and a norepinephrine transporter, can detect the change of the heart sympathetic nerve at early stage, and is the early stage of heart diseasesDiagnosis and treatment provide strong evidence; the preparation method has good repeatability and stable synthesis yield.)

1. An N- (2-fluoro [18F ] ethyl) dopamine imaging agent characterized by: the chemical structural formula of the developer is shown as the formula (I):

。

2. the method of preparing an N- (2-fluoro [18F ] ethyl) dopamine imaging agent according to claim 1, characterized in that:

the preparation method specifically comprises the following steps:

(1) transferring the fluoride ion (18F-) produced by the cyclotron into a chemical synthesizer to be captured on an anion capture column (QMA), eluting 18F-with a K2CO3/K2.2.2 solution to a reaction tube, and adding anhydrous acetonitrile to remove water azeotropically;

(2) adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) to react at 85-105 ℃ for 5-10min, and cooling the reaction solution to 60 ℃;

；

(3) adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with structural formula shown in formula (III), reacting at 80-90 deg.C for 5-10min, and cooling the reaction solution to 40 deg.C;

；

(4) injecting the reaction liquid into a semi-preparative High Performance Liquid Chromatograph (HPLC) for separation and purification, and collecting the required radioactive component, wherein the radioactive component is N- (2-fluoro [18F ] ethyl) dopamine shown in a formula (I).

3. The method of preparing an N- (2-fluoro [18F ] ethyl) dopamine imaging agent according to claim 2, characterized in that:

the preparation method specifically comprises the following steps:

(1) preparation of fluorine-18 ions

Preparing fluorinion (18F-) through a nuclear reaction of 18O (p, n) 18F by a cyclotron, transferring target water containing 18F < - > into a chemical synthesizer, then capturing the 18F < - > on an anion capture column (QMA), rinsing the QMA column by using 1.5mL of K2CO3/K2.2.2 solution, enabling the 18F < - > to enter a reaction tube, adding dry acetonitrile, and heating the reaction tube to remove water azeotropically.

4. The method of preparing an N- (2-fluoro [18F ] ethyl) dopamine imaging agent according to claim 2, characterized in that:

the preparation method specifically comprises the following steps:

(2) synthesis of N- (2-fluoro [18F ] ethyl) dopamine

Adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) into a reaction tube, heating the reaction tube to react at 85-105 ℃ for 10min, then cooling the reaction liquid to 60 ℃, adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with the structural formula shown in formula (III) into the reaction tube to react at 80-90 ℃ for 5-10min, and then cooling the reaction liquid to 40 ℃.

5. The method for preparing an N- (2-fluoro [18F ] ethyl) dopamine imaging agent according to claim 2 or 3 or 4, characterized in that:

the preparation method specifically comprises the following steps:

(3) high performance liquid chromatography separation and purification of N- (2-fluoro [18F ] ethyl) dopamine

Injecting the reaction solution into a semi-preparative High Performance Liquid Chromatograph (HPLC) to perform separation and purification on a reversed phase C18 column, wherein HPLC conditions are as follows: the mobile phase is 2% ethanol solution containing 20mM NaH2PO4, the flow rate is 5 mL/min, and the ultraviolet wavelength is set to be 220 nm; collecting radioactive components for 8-10 min, wherein the radioactive components are N- (2-fluoro [18F ] ethyl) dopamine with a structural formula shown in a formula (I), the components are filtered by a sterile filter membrane to obtain an imaging agent N- (2-fluoro [18F ] ethyl) dopamine, and the N- (2-fluoro [18F ] ethyl) dopamine imaging agent is a colorless aqueous solution containing 2% of ethanol.

Technical Field

The invention belongs to the technical field of positron nuclide imaging agents, relates to a labeling technology of radioactive compounds (preparations containing radioactive substances), and particularly relates to an N- (2-fluoro [18F ] ethyl) dopamine imaging agent and a preparation method thereof.

Background

Normal cardiovascular function depends on cardiac innervation. The adaptation of the heart to the hemodynamic requirements of the body in physiological and pathological states, in particular the rhythm, conduction and contractility of the heart, is largely regulated by the Automatic Nervous System (ANS). In various heart diseases (such as acute and chronic myocardial ischemia, myocardial infarction, arrhythmia, hypertension, diabetes, hypertrophic cardiomyopathy, familial autonomic dysfunction and the like), the change of the ANS of the heart occurs before the heart has obvious structural and functional abnormalities, and cannot be observed by a conventional morphological and functional detection method. Therefore, establishing an evaluation method of heart ANS change has important significance for early diagnosis of diseases. With the development of radioactive tracing technology and the increasing maturity of molecular imaging technology, the non-invasive methods of single photon emission tomography (SPECT) and Positron Emission Tomography (PET) can evaluate the function of the heart ANS, and further study the pathophysiological processes of nerve endings, synaptic clefts and postsynaptic receptors of the heart ANS and acquire pathophysiological information of the heart ANS in a disease state. The imaging agent is the premise of imaging, and various cardiac sympathetic nerve imaging agents clinically used at present are classified into presynaptic and postsynaptic function imaging agents according to the action parts of the imaging agents; the method is divided into SPECT and PET cardiac nerve imaging according to imaging instruments.

PET cardiac sympathetic nerve imaging agents are classified into presynaptic and postsynaptic functional imaging agents by the site of action of the imaging agent:

(1) postsynaptic imaging agents, such agents binding primarily to receptors of the postsynaptic membrane, primarily reflecting the postsynaptic adrenergic receptor distribution. Including beta adrenergic receptor (11C-CGP-12177, 11C-Carazolol) and alpha adrenergic receptor (e-GB67)) imaging agents. Representative imaging agent 11C-CGP-12177 is a hydrophilic receptor antagonist with high affinity for the β adrenergic receptor. CGP-12177 is capable of selectively recognizing beta adrenergic receptors coupled to adenylate cyclase at the cell surface. The method is mainly applied to the evaluation of the down regulation of the congenital ventricular tachycardia and the ventricular fibrillation beta-adrenergic receptor density under the condition of the dilated cardiomyopathy heart failure. But because the compound is used as precursor chlorine carbonate (phosgene), the synthesis condition is more severe, and the clinical routine application has certain difficulty;

(2) presynaptic function imaging agents including catecholamines (e.g., dopamine, norepinephrine, epinephrine) and catecholamine analogs (levo-metahydroxyephedrine). Catecholamine analogues, i.e., pseudoneurotransmitters, have good metabolic stability and low affinity for postsynaptic membrane receptors, and are not physiologically active. Including derivatives of Norepinephrine (NE), such as Metahydroxylamine (MR), metahydroxyephedrine (HED), Phenylephrine (PHEN), and antihypertensive drug (piperidine), such as Metaiodobenzylguanidine (MIBG) and its analogs. Can reflect the uptake and storage of catecholamine. A representative imaging agent of this class, 11C-HED, is taken up by sympathetic nerve endings through the same mechanism as norepinephrine-uptake-1, but is not degraded by monoamine oxidase and catechol methyltransferase as norepinephrine does during myocardial metabolism. The 11C-HED imaging can directly reflect the distribution of adrenergic receptors in organs, and is often used for clinical imaging of congestive heart failure diagnosis, heart transplantation detection, arrhythmia, diabetic autonomic neuropathy and the like. However, it has been considered that the storage and uptake processes are not completely consistent with physiological neurotransmitters.

Catecholamines include NE, Epinephrine (EPI) and Dopamine (DA), which reflect catecholamine uptake and storage:

1) NE class: the uptake mechanism, vesicle storage and metabolism of 11C-NE are more normal physiological, and therefore more suitable for evaluating the presynaptic function of sympathetic nerves. After F is introduced at the 6-position of the aromatic ring of NE (FNE), the biological process before synapse is not significantly different from NE. The clearance rate of 18F-FNE in the heart is high, so that the 18F-FNE is easily metabolized by MAO in cytoplasm, and therefore, the 18F-FNE can better reflect the in vivo process of endogenous NE. At present, 11C-NE is applied to myocardial imaging after heart in-situ transplantation, and the recovery condition of cardiac innervation can be observed;

2) epinephrine (Epinephrine EPI): the selective uptake and retention mechanism of 11C-EPI is similar to that of NE. It is metabolized by MAO in the cytosol, but its radioactive metabolites are not retained in the cell, and therefore better reflects the function of storage vesicles in the sympathetic nerve than pseudoneurotransmitter-based imaging agents that are not metabolized by MAO. It has been shown that 11C-EPI is more sensitive to detect sympathetic neuropathy than 11C-HED (pseudoneurotransmitter);

3) class DA: the studies show that the catecholamine imaging agent has higher affinity than the catecholamine analogue imaging agent receptor, and also in the catecholamine imaging agent, the dopamine imaging agent has higher affinity than the norepinephrine transporter, the mie constant of the dopamine imaging agent is 0.14 mu mol/L, and the mie constant of the dopamine imaging agent is only 0.46 mu mol/L. Fluorinated DA (FDA) can be synthesized by fluorinated substitution of the 2, 5, and 6 positions on the DA aromatic ring, respectively, and 6-fluoro-dopamine (6-18F-Fluorodopamine, 6-F-FDA) is the most studied. FDA is similar to DA biological processes in vivo, with its uptake and retention in the heart being predominantly transported via NET, followed by β -hydroxylation within vesicles, converting to fluorinated NE. At present 6-fluoro-dopamine is the imaging agent most used for clinical evaluation of primary and secondary cardiac neuropathy except 123I-MIBG. However, the synthesis of 6-18F-Fluorodopamine requires special equipment, so that the electrophilic synthesis has the problems of high cost, high synthesis difficulty, low yield and the like, and is not generally applied.

Disclosure of Invention

In view of the above, an object of the present invention is to provide a labeling compound which is more convenient and practical in a labeling method: n- (2-fluoro [18F ] ethyl) dopamine imaging agents for radionuclide imaging with strong affinity for dopamine and norepinephrine transporters.

The second purpose of the invention is to provide a preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer, and the method can successfully prepare a target compound.

In order to achieve the purpose, the technical scheme of the invention is as follows:

an N- (2-fluoro [18F ] ethyl) dopamine imaging agent characterized by: the chemical structural formula of the developer is shown as the formula (I):

。

the preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(1) transferring the fluoride ion (18F-) produced by the cyclotron into a chemical synthesizer to be captured on an anion capture column (QMA), eluting 18F-with a K2CO3/K2.2.2 solution to a reaction tube, and adding anhydrous acetonitrile to remove water azeotropically;

(2) adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) to react at 85-105 ℃ for 5-10min, and cooling the reaction solution to 60 ℃;

；

(3) adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with structural formula shown in formula (III), reacting at 80-90 deg.C for 5-10min, and cooling the reaction solution to 40 deg.C;

；

(4) injecting the reaction liquid into a semi-preparative High Performance Liquid Chromatograph (HPLC) for separation and purification, and collecting the required radioactive component, wherein the radioactive component is N- (2-fluoro [18F ] ethyl) dopamine shown in a formula (I).

The preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(1) preparation of fluorine-18 ions

Preparing fluorinion (18F-) through a nuclear reaction of 18O (p, n) 18F by a cyclotron, transferring target water containing 18F < - > into a chemical synthesizer, then capturing the 18F < - > on an anion capture column (QMA), rinsing the QMA column by using 1.5mL of K2CO3/K2.2.2 solution, enabling the 18F < - > to enter a reaction tube, adding anhydrous acetonitrile, and heating the reaction tube to remove water azeotropically.

The preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(2) synthesis of N- (2-fluoro [18F ] ethyl) dopamine

Adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) into a reaction tube, heating the reaction tube to react at 85-105 ℃ for 10min, then cooling the reaction liquid to 60 ℃, adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with the structural formula shown in formula (III) into the reaction tube to react at 80-90 ℃ for 5-10min, and then cooling the reaction liquid to 40 ℃.

The preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(3) high performance liquid chromatography separation and purification of N- (2-fluoro [18F ] ethyl) dopamine

Injecting the reaction solution into a semi-preparative High Performance Liquid Chromatograph (HPLC) to perform separation and purification on a reversed phase C18 column, wherein HPLC conditions are as follows: the mobile phase is 2% ethanol solution containing 20mM NaH2PO4, the flow rate is 5 mL/min, and the ultraviolet wavelength is set to be 220 nm; collecting radioactive components for 8-10 min, wherein the radioactive components are N- (2-fluoro [18F ] ethyl) dopamine with a structural formula shown in a formula (I), the components are filtered by a sterile filter membrane to obtain an imaging agent N- (2-fluoro [18F ] ethyl) dopamine, and the imaging agent N- (2-fluoro [18F ] ethyl) dopamine is a colorless aqueous solution containing 2% of ethanol.

The invention has the beneficial effects that: the N- (2-fluoro [18F ] ethyl) dopamine solution (18F-N-CH 2FCH2-dopamine, 18F-FEDA) has strong affinity with dopamine transporter and norepinephrine transporter; the kit can discover the change of the heart sympathetic nerve at an early stage, and provides strong evidence for early diagnosis and treatment of heart diseases.

In the preparation process, the defects that the preparation cost of 6-18F-Fluorodopamine is high and the synthesized product has a carrier are avoided. In addition, no accelerator capable of providing F + ions required by 6-18F-Fluorodopamine production exists at home, and F + needs to be transmitted by F2, so that F2 gas has the defects of active chemical property, high danger level, need of special equipment, high price and the like. The invention overcomes the difficulties, realizes the automatic synthesis of 18F-FEDA by utilizing the existing accelerator and synthesizer in China, has good repeatability of the preparation method and stable synthesis yield, has the yield of 10 percent without calibration, can be repeatedly produced in a short time, and can be prepared by all the units capable of producing positron medicines in China.

The N- (2-fluoro [18F ] ethyl) dopamine produced by the invention is used as an imaging agent, the image quality of the N- (2-fluoro [18F ] ethyl) dopamine is clearer compared with the imaging agent image described in the background technology, and smaller focuses can be found. And the half-life of the fluorine-18 is 110min, the half-life is moderate, and the fluorine-18 can be distributed to a unit without a cyclotron for use.

Drawings

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail with reference to the accompanying drawings, in which:

FIG. 1 shows an imaging agent according to the present invention: a synthetic scheme having a structural formula as shown in formula (I);

FIG. 2 shows an imaging agent according to the present invention:¹⁸HPLC identification of F-FEDA (. lamda.)_UV=220nm)；

Wherein:

a: 1 dopamine, 2¹⁹The ultraviolet absorption peak of F-FEDA,

b：¹⁸the ultraviolet absorption peak of F-FEDA,

c：¹⁸a radioactive detection peak of F-FEDA;

FIG. 3 is a drawing showing¹⁹Of F-FEDA¹H NMR mass spectrum identification chart;

FIG. 4 is a drawing showing ¹⁹Of F-FEDA¹⁹F NMR identification chart by mass spectrum.

Detailed Description

Example 1: mono, N- (2-fluoro [18F ] ethyl) dopamine developer and preparation method thereof

An N- (2-fluoro [18F ] ethyl) dopamine imaging agent characterized by: the chemical structural formula of the developer is shown as the formula (I):

。

the preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(1) transferring the fluorine ions (18F-) produced by the cyclotron into a chemical synthesizer to be captured on an anion capture column (QMA), eluting 18F-with a K2CO3/K2.2.2 solution to a reaction tube, and adding dry acetonitrile to remove water azeotropically;

(2) adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) to react at 85-105 ℃ for 5-10min, and cooling the reaction solution to 60 ℃;

；

(3) adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with structural formula shown in formula (III), reacting at 80-90 deg.C for 5-10min, and cooling the reaction solution to 40 deg.C;

；

(4) injecting the reaction liquid into a semi-preparative High Performance Liquid Chromatograph (HPLC) for separation and purification, and collecting the required radioactive component, wherein the radioactive component is N- (2-fluoro [18F ] ethyl) dopamine shown in a formula (I).

The preparation method of the N- (2-fluoro [18F ] ethyl) dopamine developer is characterized by comprising the following steps: the preparation method specifically comprises the following steps:

(1) preparation of fluorine-18 ions

Preparing fluorinion (18F-) through a nuclear reaction of 18O (p, n) 18F by a cyclotron, transferring target water containing 18F < - > into a chemical synthesizer, then capturing the 18F < - > on an anion capture column (QMA), rinsing the QMA column by using 1.5mL of K2CO3/K2.2.2 solution, enabling the 18F < - > to enter a reaction tube, adding anhydrous acetonitrile, and heating the reaction tube to remove water azeotropically.

(2) Synthesis of N- (2-fluoro [18F ] ethyl) dopamine

Adding 1mL acetonitrile solution containing 5-10mg 1, 2-bis (p-toluenesulfonyloxy) ethane with the structural formula shown in formula (II) into a reaction tube, heating the reaction tube to react at 85-105 ℃ for 5-10min, then cooling the reaction liquid to 60 ℃, adding 1mL dimethyl sulfoxide solution containing 5-10mg dopamine with the structural formula shown in formula (III) to react at 80-90 ℃ for 5-10min, and then cooling the reaction liquid to 40 ℃.

(3) High performance liquid chromatography separation and purification of N- (2-fluoro [18F ] ethyl) dopamine

Injecting the reaction solution into a semi-preparative High Performance Liquid Chromatograph (HPLC) to perform separation and purification on a reversed phase C18 column, wherein HPLC conditions are as follows: the mobile phase is 2% ethanol solution containing 20mM NaH2PO4, the flow rate is 5 mL/min, and the ultraviolet wavelength is set to be 220 nm; collecting radioactive components for 8-10 min, wherein the radioactive components are N- (2-fluoro [18F ] ethyl) dopamine with a structural formula shown in a formula (I), the components are filtered by a sterile filter membrane to obtain an imaging agent N- (2-fluoro [18F ] ethyl) dopamine, and the imaging agent N- (2-fluoro [18F ] ethyl) dopamine is a colorless aqueous solution containing 2% of ethanol.

The N- (2-fluoro [18F ] ethyl) dopamine solution (18F-N-CH 2FCH2-dopamine, 18F-FEDA) has strong affinity with dopamine transporter and norepinephrine transporter; the kit can discover the change of the heart sympathetic nerve at an early stage, and provides strong evidence for early diagnosis and treatment of heart diseases.

In the preparation process, the defects that the preparation cost of 6-18F-Fluorodopamine is high and the synthesized product has a carrier are avoided. In addition, no accelerator capable of providing F + ions required by 6-18F-Fluorodopamine production exists at home, and F + needs to be transmitted by F2, so that F2 gas has the defects of active chemical property, high danger level, need of special equipment, high price and the like. The invention overcomes the difficulties, realizes the automatic synthesis of 18F-FEDA by utilizing the existing accelerator and synthesizer in China, has good repeatability of the preparation method and stable synthesis yield, has the yield of 10 percent without calibration, can be repeatedly produced in a short time, and can be prepared by all the units capable of producing positron medicines in China.

The N- (2-fluoro [18F ] ethyl) dopamine produced by the invention is used as an imaging agent, the image quality of the N- (2-fluoro [18F ] ethyl) dopamine is clearer compared with the imaging agent image described in the background technology, and smaller focuses can be found. And the half-life of the fluorine-18 is 110min, the half-life is moderate, and the fluorine-18 can be distributed to a unit without a cyclotron for use.

Example 2: II,¹⁸Biodistribution of F-FEDA

¹⁸The biological distribution characteristics of F-FEDA meet the requirements of being used as an imaging agent, and are explained by organ distribution experiments in mice.

Taking 30 Kunming mice, half of each male and female, randomly dividing into 5 groups, each group comprising 6 mice, and injecting filtrate filtered by sterile filter membrane, i.e. the product¹⁸F-FEDA 7.4MBq (volume is less than or equal to 0.2mL), a group of animals are killed at 5 time points of 2 min, 10min, 20 min, 30 min and 60min, 11 main organs such as heart, lung, liver, spleen, kidney, stomach, intestine, brain, muscle, bone, blood and the like are taken out, weighing is carried out, radioactivity counting is carried out by a gamma counter, and percent injection dosage rate per gram of tissue (% ID/g) and radioactivity counting per gram of tissue or organ/total radioactivity counting injected into mice is multiplied by 100 percent are calculated.

The experimental results show that: heart pair¹⁸F-FEDA has very high intake, relatively little intake of lung, very high ratio of body to target and highest count of liver and spleen, which proves that¹⁸F-FEDA is metabolized by the liver.

TABLE 1,¹⁸Biodistribution of F-FEDA in Kunming species mice

。

Finally, it is noted that the above-mentioned embodiments illustrate rather than limit the invention, and that, while the invention has been described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.