CN101555232B - Pyridazinone compound marked by fluorine-18, preparation method and applications - Google Patents

Abstract

The invention discloses a pyridazinone compound marked by fluorine-18 with a molecular formula of <18>FFPnOP and a preparation method and applications; in the formula, n is equal to 1, 2 or 3. By technical synthesis to ligand OTs-PnOP, the pyridazinone compound <18>FFPnOP marked by radioactive fluorine-18 and stable reference compound <19>FFPnOP are obtained by synthesis, wherein the stable reference compound is used for confirming the structure of the compound with radioactive marks; the compound has high radiochemical purity, good biological properties, high initial uptake value, simple preparation and low use cost, and is applied in the technical fields of radioactive drug chemistry and clinical nuclear medicine as a novel myocardial perfusion imaging agent marked by fluorine-18.

Description

Translated fromChinese

氟-18标记哒嗪酮类化合物及制备方法和应用Fluorine-18 labeled pyridazinone compounds, preparation method and application

所属技术领域Technical field

本发明涉及到氟-18标记的放射性药物化学和临床核医学技术领域，特别是涉及到一类氟-18标记哒嗪酮类化合物及制备方法和应用。The invention relates to the technical fields of fluorine-18 labeled radiopharmaceutical chemistry and clinical nuclear medicine, in particular to a class of fluorine-18 labeled pyridazinone compounds, their preparation method and application.

背景技术Background technique

冠心病是威胁人类健康最严重的疾病之一。在我国，冠心病的发病率以及死亡率均呈上升趋势。2007年，卫生部公布的资料显示：我国每年死于心血管疾病的约300万人，已成为我国城乡人群的第一位死亡原因，占我国居民死亡原因的近40％。Coronary heart disease is one of the most serious diseases threatening human health. In my country, the morbidity and mortality of coronary heart disease are on the rise. In 2007, the data released by the Ministry of Health showed that about 3 million people died of cardiovascular disease in my country every year, which has become the first cause of death of urban and rural population in my country, accounting for nearly 40% of the causes of death of Chinese residents.

心肌灌注显像用于心脏病无创检查始于上世纪70年代，其巨大的诊断价值已在世界范围内被广泛接受，成为目前冠心病诊断、疗效评价以及预后判断的最重要的影像学方法之一。心肌灌注单光子发射计算机断层(SPECT)显像技术是当前临床上用于冠心病检测的无创性灌注显像的主要方法。但是，与SPECT相比，正电子发射断层(PET)显像有更高的空间和时间分辨率，可以有效降低组织衰减，利用标准的组织衰减校正方法可以做到冠状血流的绝对定量。另外，正电子核素的短半衰期可有效减少靶组织周围的辐射剂量，短半衰期也能缩短静息和运动显像间隔。常用的心肌灌注PET显像剂包括：¹⁵O-H₂O，¹³N-NH₃和⁸²Rb等。但上述显像剂半衰期均较短，临床应用还受到很大的限制。而¹⁸F相对于其他正电子核素，半衰期较长(t_1/2＝109.8min)；具有较低的正电子能量(512keV)，对正常组织的辐射损伤较小；其范德华半径(1.35

)与氢(1.2

)相似，不会影响标记化合物的生物活性，因此，研制新型的氟-18标记的心肌灌注显像剂具有重要的现实意义。The use of myocardial perfusion imaging in non-invasive heart disease examinations began in the 1970s, and its great diagnostic value has been widely accepted around the world. It has become one of the most important imaging methods for the diagnosis, efficacy evaluation and prognosis of coronary heart disease. one. Myocardial perfusion single photon emission computed tomography (SPECT) imaging technology is currently the main method of non-invasive perfusion imaging clinically used in the detection of coronary heart disease. However, compared with SPECT, positron emission tomography (PET) imaging has higher spatial and temporal resolution, can effectively reduce tissue attenuation, and can achieve absolute quantification of coronary blood flow using standard tissue attenuation correction methods. In addition, the short half-life of positron nuclides can effectively reduce the radiation dose around the target tissue, and the short half-life can also shorten the interval between resting and motion imaging. Commonly used myocardial perfusion PET imaging agents include:¹⁵ OH₂ O,¹³ N-NH₃ and⁸² Rb. However, the half-life of the above-mentioned imaging agents is short, and the clinical application is still greatly limited. Compared with other positron nuclides,¹⁸ F has a longer half-life (t_1/2 = 109.8min); has a lower positron energy (512keV), and has less radiation damage to normal tissues; its van der Waals radius (1.35

) and hydrogen (1.2

) are similar and will not affect the biological activity of the labeled compound. Therefore, the development of a new type of fluorine-18 labeled myocardial perfusion imaging agent has important practical significance.

心肌组织中，线粒体的重量约占30％。在绝大多数的哺乳动物组织中，80％～90％的ATP产生于线粒体内的氧化磷酸化过程。该过程由位于线粒体内膜中的呼吸链(电子传递链)完成。研究表明：线粒体呼吸链由四种线粒体复合体(MC-I、II、III、IV)组成，其中MC-I(NADH-泛醌氧化还原酶)是该链的开端。MC-I是一种有40多个不同亚单位的膜结合蛋白，是电子的进入位点。最近报道的几种PET心肌灌注显像剂均为MC-I抑制剂类似物，如¹⁸F-FDHR，¹⁸F-4-(1，3-二氮杂萘衍生物)，BMS-747158-02等。哒螨灵是MC-I的抑制剂之一，如何对哒螨灵进行结构修饰，将其转化为可用于氟-18标记的哒嗪酮类标记前体，并制备氟-18标记的哒嗪酮类心肌灌注显像剂是本技术领域需要解决的课题。In myocardial tissue, mitochondria account for about 30% by weight. In most mammalian tissues, 80% to 90% of ATP is produced in the process of oxidative phosphorylation in mitochondria. This process is accomplished by the respiratory chain (electron transport chain) located in the inner membrane of the mitochondria. Studies have shown that the mitochondrial respiratory chain consists of four mitochondrial complexes (MC-I, II, III, IV), among which MC-I (NADH-ubiquinone oxidoreductase) is the beginning of the chain. MC-I is a membrane-bound protein with more than 40 different subunits, which are the entry sites for electrons. Several recently reported PET myocardial perfusion imaging agents are MC-I inhibitor analogs, such as¹⁸ F-FDHR,¹⁸ F-4-(1,3-naphthyridine derivatives), BMS-747158-02 wait. Pyridaben is one of the inhibitors of MC-I. How to modify the structure of pyridaben to convert it into a pyridazinone-labeled precursor that can be used for fluorine-18 labeling, and prepare fluorine-18-labeled pyridazine The ketone myocardial perfusion imaging agent is a subject to be solved in this technical field.

发明内容Contents of the invention

本发明的目的是提供一类放射化学纯度高、生物性能好、初始摄取值高、制备简单及使用成本低，应用在心肌灌注正电子发射型计算机断层显像领域的氟-18标记的新哒嗪酮类([¹⁸F]FPnOP)化合物。The purpose of the present invention is to provide a new class of fluorine-18-labeled fluorine-18 with high radiochemical purity, good biological performance, high initial uptake value, simple preparation and low use cost, which can be used in the field of myocardial perfusion positron emission computed tomography imaging. Azinones ([¹⁸ F]FPnOP) compounds.

为了达到上述目的，本发明采用以下技术方案：一类氟-18标记的哒嗪酮类化合物，其分子式为[¹⁸F]FPnOP，其结构通式如下：In order to achieve the above object, the present invention adopts the following technical scheme: a class of pyridazinone compounds labeled with fluorine-18, the molecular formula of which is [¹⁸ F]FPnOP, and its general structural formula is as follows:

其中，n＝1、2、3。Among them, n=1, 2, 3.

氟-18标记的哒嗪酮类化合物的制备方法如下：The preparation method of fluorine-18 labeled pyridazinone compounds is as follows:

(1)配体OTs-PnOP的合成：(1) Synthesis of ligand OTs-PnOP:

a.冰浴下，将适量MnO₂和糠醛分批加至适量浓盐酸中，20～100℃反应1～2h，冷却过夜，抽滤，重结晶得到糠氯酸(DCA)。a. Under an ice bath, add an appropriate amount of MnO₂ and furfural to an appropriate amount of concentrated hydrochloric acid in batches, react at 20-100°C for 1-2 hours, cool overnight, filter with suction, and recrystallize to obtain furochloric acid (DCA).

b.冰浴下，将适量糠氯酸、Na₂CO₃和叔丁基肼盐酸盐加至适量水中，搅拌1～3h，抽滤。将沉淀加至适量苯和冰醋酸中，10～60℃反应3～6h，得到4，5-二氯哒嗪酮(DCP)。b. Under ice bath, add appropriate amount of furochloric acid, Na₂ CO₃ and tert-butylhydrazine hydrochloride into appropriate amount of water, stir for 1-3 hours, and filter with suction. Add the precipitate to an appropriate amount of benzene and glacial acetic acid, and react at 10-60°C for 3-6 hours to obtain 4,5-dichloropyridazinone (DCP).

c.将适量4-羟基苯甲酸甲酯、K₂CO₃、KI和PEGn(n＝1～3)悬浮于适量环己酮中，回流12-24h，得到MB-PEGn(n＝1～3)。PEGn：当n＝1时为2-氯乙醇；当n＝2时为2-2[-氯乙氧基]乙醇；当n＝3时为2-2[-2[-氯乙氧基]乙氧基]乙醇。c. Suspend an appropriate amount of methyl 4-hydroxybenzoate, K₂ CO₃ , KI and PEGn (n=1~3) in an appropriate amount of cyclohexanone, and reflux for 12-24 hours to obtain MB-PEGn (n=1~3 ). PEGn: 2-chloroethanol when n=1; 2-2[-chloroethoxy]ethanol when n=2; 2-2[-2[-chloroethoxy] when n=3 Ethoxy]ethanol.

d.将适量MB-PEGn(n＝1～3)、咪唑、二甲基叔丁基氯硅烷加至适量DMF中，50～100℃反应1～5h，得到MSB-PEGn(n＝1～3)。d. Add an appropriate amount of MB-PEGn (n=1~3), imidazole, and dimethyl tert-butylchlorosilane to an appropriate amount of DMF, and react at 50~100°C for 1~5h to obtain MSB-PEGn (n=1~3 ).

e.冰浴下，将适量MSB-PEGn(n＝1～3)、氢化锂铝加至适量无水乙醚中，0～50℃搅拌2～3h，得到SPE-PEGn(n＝1～3)。e. Under an ice bath, add an appropriate amount of MSB-PEGn (n=1~3) and lithium aluminum hydride to an appropriate amount of anhydrous ether, and stir at 0~50°C for 2~3 hours to obtain SPE-PEGn (n=1~3) .

f.将适量4，5-二氯哒嗪酮、碳酸铯、SPE-PEGn(n＝1～3)加至适量无水DMF中，70～100℃反应12～24h，得到SB-PnOP(n＝1～3)。f. Add an appropriate amount of 4,5-dichloropyridazinone, cesium carbonate, and SPE-PEGn (n=1 to 3) to an appropriate amount of anhydrous DMF, and react at 70 to 100°C for 12 to 24 hours to obtain SB-PnOP (n =1~3).

g.将适量1mol/L四正丁基氟化铵(TBAF)的四氢呋喃溶液和SB-PnOP(n＝1～3)加至适量四氢呋喃中，10～70℃反应1～2h，得到HB-PnOP(n＝1～3)。g. Add an appropriate amount of 1mol/L tetrahydrofuran solution of tetra-n-butylammonium fluoride (TBAF) and SB-PnOP (n=1~3) to an appropriate amount of tetrahydrofuran, and react at 10~70°C for 1~2h to obtain HB-PnOP (n = 1 ~ 3).

h.将适量对甲基苯磺酰氯、4-二甲氨基吡啶、二异丙基乙胺、HB-PnOP(n＝1～3)加至无水CH₂Cl₂中，5～60℃反应2～4h，得到OTs-PnOP(n＝1～3)。其合成路线为：h. Add appropriate amount of p-toluenesulfonyl chloride, 4-dimethylaminopyridine, diisopropylethylamine, HB-PnOP (n=1~3) to anhydrous CH₂ Cl₂ and react at 5~60°C After 2-4 hours, OTs-PnOP (n=1-3) was obtained. Its synthetic route is:

(2)[¹⁹F]FPnOP(n＝1～3)的合成(2) Synthesis of [¹⁹ F]FPnOP (n=1-3)

将适量OTs-PnOP(n＝1～3)和四正丁基氟化铵(TBAF)加至适量无水乙腈中，70～100℃反应20～60min，得到[¹⁹F]FPnOP(n＝1～3)。Add an appropriate amount of OTs-PnOP (n=1~3) and tetra-n-butylammonium fluoride (TBAF) to an appropriate amount of anhydrous acetonitrile, and react at 70~100°C for 20~60min to obtain [¹⁹ F]FPnOP (n=1 ~3).

其合成路线为：Its synthetic route is:

(3)氟-18标记的哒嗪酮类化合物([¹⁸F]FPnOP，n＝1～3)的合成(3) Synthesis of fluorine-18-labeled pyridazinones ([¹⁸ F]FPnOP, n=1-3)

将K₂₂₂、K₂CO₃、¹⁸[F]F^-和OTs-PnOP(n＝1～3)加至适量无水乙腈中，70～100℃反应20～60min，得到[¹⁸F]FPnOP(n＝1～3)。Add K₂₂₂ , K₂ CO₃ ,¹⁸ [F]F^- and OTs-PnOP (n=1~3) to an appropriate amount of anhydrous acetonitrile, and react at 70~100°C for 20~60min to obtain [¹⁸ F]FPnOP ( n=1~3).

其合成路线为：Its synthetic route is:

上述所述的化学合成试剂均是市售商品，来源广泛，容易获得。The chemical synthesis reagents mentioned above are all commercially available, widely sourced and easy to obtain.

本发明以放射性¹⁸[F]F^-和OTs-PnOP(n＝1～3)反应制备得到所述的放射性氟-18标记的哒嗪酮类化合物([¹⁸F]FPnOP，n＝1～3)，是一种新型的心肌灌注显像剂。[¹⁸F]FPnOP(n＝1～3)分子中放射性的氟-18，可用于正电子发射型计算机断层显像(PET)。[¹⁸F]FPnOP(n＝1～3)的放射化学纯度高、生物性能好、初始摄取值及靶与非靶比高。与已报道的正电子心肌灌注显像剂相比，其生物性能更佳，可以成为一种新型的心肌灌注显像剂在临床上应用推广。The present invention prepares the radioactive fluorine-18-labeled pyridazinone compound ([¹⁸ F]FPnOP, n=1-3) by reacting radioactive¹⁸ [F^] F- and OTs-PnOP (n=1-3). ), is a new type of myocardial perfusion imaging agent. The radioactive fluorine-18 in [¹⁸ F]FPnOP (n=1-3) molecules can be used in positron emission computed tomography (PET). [¹⁸ F]FPnOP (n=1-3) has high radiochemical purity, good biological performance, high initial uptake value and high ratio of target to non-target. Compared with the reported positron myocardial perfusion imaging agent, its biological performance is better, and it can become a new type of myocardial perfusion imaging agent for clinical application and promotion.

以[¹⁸F]FP2OP为例，其衰变校正后的放化产率为30～55％，放化纯度＞99％。Taking [¹⁸ F]FP2OP as an example, its decay-corrected radiochemical yield is 30-55%, and its radiochemical purity is >99%.

实验表明，[¹⁸F]FP2OP的基本的性能如下：Experiments show that the basic performance of [¹⁸ F]FP2OP is as follows:

1.[¹⁸F]FP2OP在正常小鼠体内生物分布1. Biodistribution of [¹⁸ F]FP2OP in normal mice

取20只正常昆明小白鼠，于尾静脉注射0.1mL[¹⁸F]FP2OP(约0.296MBq)。于注射后2、15、30、60min后断头处死。取出心、肝、肺、肾、肌肉、骨、血等组织，称量并在γ计数器中测其放射性计数。[¹⁸F]FP2OP在正常小鼠中的生物分布见表1。20 normal Kunming mice were injected with 0.1 mL [¹⁸ F]FP2OP (about 0.296 MBq) in the tail vein. They were decapitated 2, 15, 30, and 60 minutes after injection. Take out heart, liver, lung, kidney, muscle, bone, blood and other tissues, weigh them and measure their radioactive counts in a gamma counter. The biodistribution of [¹⁸ F]FP2OP in normal mice is shown in Table 1.

表1.[¹⁸F]FP2OP在小鼠体内的生物分布(％ID/g±SD，n＝5)Table 1. Biodistribution of [¹⁸ F]FP2OP in mice (%ID/g±SD, n=5)

在正常小鼠中的生物分布结果显示，[¹⁸F]FP2OP在心肌中有很高的初始摄取且有一定的滞留。注射2min后，心肌摄取值为(41.90±4.52)％ID/g，注射60min后，心肌摄取仍有(24.80±0.71)％ID/g。同时，[¹⁸F]FP2OP靶与非靶比值较高，如2min时心肝比为6.83，心肺比为9.49，心血比为35.74。无论其在心肌摄取值，还是靶非靶比值方面，[¹⁸F]FP2OP均远远优于当前在临床广泛使用的心肌灌注显像剂^99mTc-MIBI，据此我们认为该类配合物有望发展为一类新型的心肌灌注显像剂。The results of biodistribution in normal mice showed that [¹⁸ F]FP2OP had a high initial uptake and a certain retention in the myocardium. After 2 minutes of injection, the myocardial uptake was (41.90±4.52)%ID/g, and after 60min of injection, the myocardial uptake was still (24.80±0.71)%ID/g. At the same time, the [¹⁸ F]FP2OP target-to-non-target ratio was high, for example, the heart-to-liver ratio was 6.83, the heart-lung ratio was 9.49, and the heart-to-blood ratio was 35.74 at 2 minutes. Regardless of its myocardial uptake value or target-to-target ratio, [¹⁸ F]FP2OP is far superior to^99m Tc-MIBI, a myocardial perfusion imaging agent widely used in clinical practice. Therefore, we believe that this type of complex is expected to develop It is a new type of myocardial perfusion imaging agent.

具体实施方式：Detailed ways:

下面以n＝2时为例，通过实施例详述本发明：一类氟-18标记的哒嗪酮类化合物：Taking n=2 as an example below, the present invention is described in detail through examples: a class of fluorine-18 labeled pyridazinone compounds:

(1)配体OTs-P2OP的合成：(1) Synthesis of ligand OTs-P2OP:

a.糠氯酸(DCA)的合成a. Synthesis of furochloric acid (DCA)

冰浴下，在1L四口瓶中加入480mL浓盐酸。0-10℃时，分批加入80g MnO₂，搅拌，缓慢滴加24g糠醛。室温搅拌30min后，缓慢升温至60℃。60～80℃时，分批加入42g MnO₂，升温至100℃。反应至液体变为橙色透明溶液后，继续反应30min，冷却，抽滤。沉淀用40mL乙醚溶解，过滤。滤液经旋蒸除去溶剂，用40mL水重结晶，得到31g糠氯酸，为浅粉色片状晶体。核磁谱图：(¹HNMR，CDCl₃)δ：6.085(s，1H，HO-C-H)；(¹³CNMR，CDCl₃)δ：98.56，123.87，151.25，165.15。红外谱图：(IR)/cm^-1：vOH：3407，vC＝O：1770，vC＝C：1638。Under ice bath, add 480mL of concentrated hydrochloric acid into a 1L four-neck flask. At 0-10°C, add 80g of MnO₂ in batches, stir, and slowly add 24g of furfural dropwise. After stirring at room temperature for 30 min, the temperature was slowly raised to 60°C. At 60-80°C, add 42g of MnO₂ in batches and raise the temperature to 100°C. After the reaction until the liquid turned into an orange transparent solution, the reaction was continued for 30 min, cooled, and suction filtered. The precipitate was dissolved with 40 mL of ether and filtered. The filtrate was evaporated to remove the solvent, and recrystallized with 40 mL of water to obtain 31 g of furochloric acid as light pink flaky crystals. NMR spectrum:^{( 1} HNMR, CDCl₃ ) δ: 6.085 (s, 1H, HO-CH); (¹³ CNMR, CDCl₃ ) δ: 98.56, 123.87, 151.25, 165.15. Infrared spectrum: (IR)/cm^-1 : vOH: 3407, vC=O: 1770, vC=C: 1638.

b.4，5-二氯哒嗪酮(DCP)的合成b. Synthesis of 4,5-dichloropyridazinone (DCP)

冰浴下，将15g糠氯酸溶于150mL水中，加入4.5g无水Na₂CO₃，搅拌至溶液变澄清。加入10g叔丁基肼盐酸盐，搅拌2.5h，抽滤。沉淀用冷水洗涤后，转移至100mL茄形瓶。加入50mL苯和7g冰醋酸，加热至35～45℃，反应4h。加入20mL水，分出苯层，分别用20mL 5％NaOH溶液、20mL 10％HCl溶液和20mL水洗涤，有机相经MgSO₄干燥，旋蒸除去大部分溶剂后静置过夜。析出6.9g 4，5-二氯哒嗪酮，为浅黄色针状晶体。核磁谱图：(¹HNMR，CDCl₃)δ：1.648(s，9H，N(CH₃)₃)，7.729(s，1H，N＝C-H)。红外谱图：(IR)/cm^-1：vOH：3461，vC＝O：1658。Under ice bath, dissolve 15g of furochloric acid in 150mL of water, add 4.5g of anhydrous Na₂ CO₃ , and stir until the solution becomes clear. Add 10g of tert-butylhydrazine hydrochloride, stir for 2.5h, and filter with suction. After the precipitate was washed with cold water, it was transferred to a 100 mL eggplant-shaped flask. Add 50mL of benzene and 7g of glacial acetic acid, heat to 35-45°C, and react for 4h. Add 20mL of water, separate the benzene layer, wash with 20mL of 5% NaOH solution, 20mL of 10% HCl solution and 20mL of water respectively, and dry the organic phase over_MgSO4 , remove most of the solvent by rotary evaporation and let it stand overnight. 6.9 g of 4,5-dichloropyridazinone was precipitated as pale yellow needle-like crystals. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 1.648 (s, 9H, N(CH₃ )₃ ), 7.729 (s, 1H, N=CH). Infrared spectrum: (IR)/cm^-1 : vOH: 3461, vC=O: 1658.

c.MB-PEG2的合成c. Synthesis of MB-PEG2

将1g 4-羟基苯甲酸甲酯、1.81g K₂CO₃和0.55g KI加至100mL三口瓶中。氮气保护下加入20mL环己酮和1.8mL 2-(2-氯乙氧基)乙醇，回流24h。冷却至室温，抽滤。滤液旋蒸后，加入15mL二氯甲烷，抽滤。滤液旋蒸后经200-300目的硅胶柱纯化，展开剂为乙醚。最后得到1.0378g MB-PEG2，为浅黄色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：2.072(s，1H，OH)，3.677(t，2H，CH₂CH₂OH)，3.769(t，2H，CH₂CH₂OH)，3.883(s，3H，OCH₃)，3.886(t，2H，phenyl-O-CH₂CH₂)，4.191(t，2H，phenyl-O-CH₂)，6.934(d，2H，O-phenyl)，7.986(d，2H，CO-phenyl)。红外谱图：(IR)/cm^-1：vOH：3461，vC＝O：1715，vC-O-C：1102。Add 1 g of methyl 4-hydroxybenzoate, 1.81 g of K₂ CO₃ and 0.55 g of KI into a 100 mL three-necked flask. Under nitrogen protection, 20 mL of cyclohexanone and 1.8 mL of 2-(2-chloroethoxy)ethanol were added, and refluxed for 24 hours. Cool to room temperature and filter with suction. After the filtrate was rotary evaporated, 15 mL of dichloromethane was added, and suction filtered. After rotary evaporation, the filtrate was purified by a 200-300 mesh silica gel column, and the developing solvent was diethyl ether. Finally, 1.0378 g of MB-PEG2 was obtained as a pale yellow oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 2.072 (s, 1H, OH), 3.677 (t, 2H, CH₂ CH₂ OH), 3.769 (t, 2H, CH₂ CH₂ OH), 3.883 ( s, 3H, OCH₃ ), 3.886 (t, 2H, phenyl-O-CH₂ CH₂ ), 4.191 (t, 2H, phenyl-O-CH₂ ), 6.934 (d, 2H, O-phenyl), 7.986 (d, 2H, CO-phenyl). Infrared spectrum: (IR)/cm^-1 : vOH: 3461, vC=O: 1715, vC-OC: 1102.

d.MSB-PEG2的合成d. Synthesis of MSB-PEG2

将1.0378g MB-PEG2、0.5023g咪唑、1.08g二甲基叔丁基氯硅烷和10mL无水DMF加至50mL三口瓶中，80℃油浴反应4h。反应液用20mL乙酸乙酯稀释后抽滤。滤液转移至100mL分液漏斗，分别用20mL水洗5次，用20mL饱和NaHCO₃溶液洗2次。有机相经MgSO₄干燥后。抽滤，旋蒸滤液得到1.0732g MSB-PEG2，为浅黄色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：0.000(s，6H，OSi(CH₃)₂)，0.824(s，9H，SiC(CH₃)₃)，3.567(t，2H，CH₂CH₂OSi)，3.726(t，2H，CH₂CH₂OSi)，3.813(s，3H，OCH₃)，3.815(t，2H，phenyl-O-CH₂CH₂)，4.099(t，2H，phenyl-O-CH₂)，6.862(d，2H，O-phenyl)，7.911(d，2H，CO-phenyl)。红外谱图：(IR)/cm^-1：vC＝O：1723，v＝C-O-C：1250，vC-O-C：1087。Add 1.0378g MB-PEG2, 0.5023g imidazole, 1.08g dimethyl tert-butylchlorosilane and 10mL anhydrous DMF to a 50mL three-necked flask, and react in an oil bath at 80°C for 4h. The reaction solution was diluted with 20 mL of ethyl acetate and filtered with suction. The filtrate was transferred to a 100mL separatory funnel, washed 5 times with 20mL water and 2 times with 20mL saturated_NaHCO3 solution. The organic phase was dried over MgSO₄ . Suction filtration and rotary evaporation of the filtrate gave 1.0732 g of MSB-PEG2 as light yellow oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 0.000(s, 6H, OSi(CH₃ )₂ ), 0.824(s, 9H, SiC(CH₃ )₃ ), 3.567(t, 2H, CH₂ CH₂ OSi), 3.726(t, 2H, CH₂ CH₂ OSi), 3.813(s, 3H, OCH₃ ), 3.815(t, 2H, phenyl-O-CH₂ CH₂ ), 4.099(t, 2H, phenyl -O-CH₂ ), 6.862 (d, 2H, O-phenyl), 7.911 (d, 2H, CO-phenyl). Infrared spectrum: (IR)/cm^-1 : vC=O: 1723, v=COC: 1250, vC-OC: 1087.

e.SPE-PEG2的合成e. Synthesis of SPE-PEG2

将0.28g LiAlH₄加至50mL三口瓶中，冰浴30min。氮气保护下，加入8mL无水乙醚。将1.0732g MBS-PEG2溶于7mL无水乙醚中，缓慢滴加至反应瓶中，反应2h。撤去冰浴，室温反应2h后，缓慢滴加乙醇和水，至无气体产生。抽滤，旋蒸滤液得到0.38g SPE-PEG，为浅黄色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：0.000(s，6H，OSi(CH₃)₂)，0.815(s，9H，SiC(CH₃)₃)，3.580(t，2H，CH₂CH₂OSi)，3.672(t，2H，CH₂CH₂OSi)，3.775(t，2H，phenyl-O-CH₂CH₂)，4.046(t，2H，phenyl-O-CH₂CH₂)，4.466(s，2H，CH₂-O-C＝C-Cl)，6.807(d，2H，O-phenyl)，7.211(d，2H，CO-phenyl)。Add 0.28g LiAlH₄ to a 50mL three-neck flask, and ice-bath for 30min. Under nitrogen protection, 8 mL of anhydrous diethyl ether was added. Dissolve 1.0732g of MBS-PEG2 in 7mL of anhydrous ether, slowly drop it into the reaction bottle, and react for 2h. Remove the ice bath, and after reacting at room temperature for 2 hours, slowly add ethanol and water dropwise until no gas is generated. Suction filtration and rotary evaporation of the filtrate yielded 0.38 g of SPE-PEG as light yellow oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 0.000(s, 6H, OSi(CH₃ )₂ ), 0.815(s, 9H, SiC(CH₃ )₃ ), 3.580(t, 2H, CH₂ CH₂ OSi), 3.672 (t, 2H, CH₂ CH₂ OSi), 3.775 (t, 2H, phenyl-O-CH₂ CH₂ ), 4.046 (t, 2H, phenyl-O-CH₂ CH₂ ), 4.466 (s, 2H, CH2_- OC=C-Cl), 6.807 (d, 2H, O-phenyl), 7.211 (d, 2H, CO-phenyl).

f.SB-P2OP的合成f. Synthesis of SB-P2OP

将0.38g SPE-PEG2、0.93g DCP、1.37g Cs₂CO₃和10mL无水DMF加至50mL茄形瓶中，68℃油浴反应12h。反应液冷却至室温，用20mL乙酸乙酯稀释后抽滤。滤液转移至100mL分液漏斗，分别用25mL水洗5次。有机相经MgSO₄干燥，抽滤后旋蒸滤液，经200-300目的硅胶柱纯化，展开剂为正己烷∶乙酸乙酯＝3∶1。最后得到0.552g SB-P2OP，为浅黄色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：0.000(s，6H，OSi(CH₃)₂)，0.824(s，9H，SiC(CH₃)₃)，1.558(s，9H，N(CH₃)₃)，3.566(t，2H，CH₂CH₂OSi)，3.727(t，2H，CH₂CH₂OSi)，3.802(t，2H，phenyl-O-CH₂CH₂)，4.057(t，2H，phenyl-OCH₂CH₂)，5.177(s，2H，CH₂-O-C＝C-Cl)，6.876(d，2H，O-phenyl)，7.255(d，2H，CO-phenyl)，7.663(s，1H，N＝C-H)。红外谱图：(IR)/cm^-1：vC＝O：1649。Add 0.38g of SPE-PEG2, 0.93g of DCP, 1.37g of Cs₂ CO₃ and 10mL of anhydrous DMF into a 50mL eggplant-shaped bottle, and react in an oil bath at 68°C for 12h. The reaction solution was cooled to room temperature, diluted with 20 mL of ethyl acetate, and filtered with suction. The filtrate was transferred to a 100mL separatory funnel and washed 5 times with 25mL water respectively. The organic phase was dried over MgSO₄ , and the filtrate was evaporated after suction filtration, and purified through a 200-300 mesh silica gel column, and the developing solvent was n-hexane:ethyl acetate=3:1. Finally, 0.552 g of SB-P2OP was obtained as a pale yellow oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 0.000(s, 6H, OSi(CH₃ )₂ ), 0.824(s, 9H, SiC(CH₃ )₃ ), 1.558(s, 9H, N(CH₃ )₃ ), 3.566(t, 2H_,CH2CH2OSi ), 3.727(t, 2H_,CH2CH2OSi ), 3.802(t, 2H, phenyl-O-_CH2CH2) , 4.057(t , 2H, phenyl-OCH₂ CH₂ ), 5.177 (s, 2H, CH₂ -OC=C-Cl), 6.876 (d, 2H, O-phenyl), 7.255 (d, 2H, CO-phenyl), 7.663 (s, 1H, N=CH). Infrared spectrum: (IR)/cm^-1 : vC=O: 1649.

g.HB-P2OP的合成g. Synthesis of HB-P2OP

将0.552g SB-P2OP、3mL 1mol/L四正丁基氟化铵(TBAF)的四氢呋喃溶液和3mL无水THF加至25mL茄形瓶中，搅拌2h。反应液用20mL乙酸乙酯稀释后，转移至100mL分液漏斗中，用20mL水洗涤。有机相用MgSO₄干燥，抽滤后旋蒸滤液，经200-300目的硅胶柱纯化，展开剂为二氯甲烷∶甲醇＝7∶1。最后得到0.361g HB-P2OP，为黄色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：1.532(s，9H，N(CH₃)₃)，3.586(t，2H，CH₂CH₂OH)，3.678(t，2H，CH₂CH₂OH)，3.785(t，2H，phenyl-O-CH₂CH₂)，4.059(t，2H，phenyl-OCH₂CH₂)，5.153(s，2H，CH₂-O-C＝C-Cl)，6.857(d，2H，O-phenyl)，7.240(d，2H，CO-phenyl)，7.636(s，1H，N＝C-H)。红外谱图：(IR)/cm^-1：vOH：3239(O-H)，vC＝O：1643，v＝C-O-C：1251，vC-O-C：1137。Add 0.552g of SB-P2OP, 3mL of 1mol/L tetrahydrofuran solution of 1mol/L tetra-n-butylammonium fluoride (TBAF) and 3mL of anhydrous THF into a 25mL eggplant-shaped flask, and stir for 2h. The reaction solution was diluted with 20 mL of ethyl acetate, transferred to a 100 mL separatory funnel, and washed with 20 mL of water. The organic phase was dried with MgSO₄ . After suction filtration, the filtrate was evaporated and purified through a 200-300 mesh silica gel column. The developing solvent was dichloromethane:methanol=7:1. Finally, 0.361 g of HB-P2OP was obtained as a yellow oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 1.532(s, 9H, N(CH₃ )₃ ), 3.586(t, 2H, CH₂ CH₂ OH), 3.678(t, 2H, CH₂ CH₂ OH), 3.785 (t, 2H, phenyl-O-CH₂ CH₂ ), 4.059 (t, 2H, phenyl-OCH₂ CH₂ ), 5.153 (s, 2H, CH₂ -OC=C-Cl), 6.857 (d, 2H, O-phenyl), 7.240 (d, 2H, CO-phenyl), 7.636 (s, 1H, N=CH). Infrared spectrum: (IR)/cm^-1 : vOH: 3239 (OH), vC=O: 1643, v=COC: 1251, vC-OC: 1137.

h.OTs-P2OP的合成Synthesis of h.OTs-P2OP

将0.361g HB-P2OP、0.252g对甲基苯磺酰氯、0.167g 4-二甲氨基吡啶、0.175g二异丙基乙胺和3mL无水二氯甲烷加至25mL茄形瓶中，搅拌2h。反应液用20mL乙酸乙酯稀释后，转移至100mL分液漏斗中，分别用20mL 0.1M HCl和20mL水洗。有机相用MgSO₄干燥，抽滤后旋蒸滤液，经200-300目的硅胶柱纯化，展开剂为二氯甲烷∶甲醇＝100∶1。最后得到OTs-P2OP，为棕色油状物。核磁谱图：(¹HNMR，CDCl₃)δ：1.558(s，9H，N(CH₃)₃)，2.341(s，3H，phenyl-CH₃)，3.695(t，2H，CH₂CH₂OS)，3.729(t，2H，CH₂CH₂OS)，3.990(t，2H，phenyl-O-CH₂CH₂)，4.122(t，2H，phenyl-OCH₂CH₂)，5.180(s，2H，CH₂-O-C＝C-Cl)，6.850(d，2H，O-phenyl)，7.235(d，2H，CO-phenyl)，7.265(d，2H，phenyl-CH₃)，7.666(s，1H，N＝C-H)，7.723(d，2H，phenyl-SO₃)。红外谱图：(IR)/cm^-1：vC＝O：1652，v＝C-O-C：1249，vC-O-C：1092。质谱：C26H31ClFN2O7S：计算值：550.1；实际值：551.2。Add 0.361g of HB-P2OP, 0.252g of p-toluenesulfonyl chloride, 0.167g of 4-dimethylaminopyridine, 0.175g of diisopropylethylamine and 3mL of anhydrous dichloromethane into a 25mL eggplant-shaped bottle and stir for 2h . The reaction solution was diluted with 20 mL of ethyl acetate, transferred to a 100 mL separatory funnel, and washed with 20 mL of 0.1M HCl and 20 mL of water, respectively. The organic phase was dried with MgSO₄ . After suction filtration, the filtrate was evaporated and purified through a 200-300 mesh silica gel column. The developing solvent was dichloromethane:methanol=100:1. Finally, OTs-P2OP was obtained as a brown oil. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 1.558(s, 9H, N(CH₃ )₃ ), 2.341(s, 3H, phenyl-CH₃ ), 3.695(t, 2H, CH₂ CH₂ OS ), 3.729 (t, 2H, CH₂ CH₂ OS), 3.990 (t, 2H, phenyl-O-CH₂ CH₂ ), 4.122 (t, 2H, phenyl-OCH₂ CH₂ ), 5.180 (s, 2H , CH₂ -OC=C-Cl), 6.850 (d, 2H, O-phenyl), 7.235 (d, 2H, CO-phenyl), 7.265 (d, 2H, phenyl-CH₃ ), 7.666 (s, 1H , N=CH), 7.723 (d, 2H, phenyl-SO₃ ). Infrared spectrum: (IR)/cm^-1 : vC=O: 1652, v=COC: 1249, vC-OC: 1092. Mass Spectrum: C26H31ClFN2O7S: Calculated: 550.1; Actual: 551.2.

(2)[¹⁹F]FP2OP的合成(2) Synthesis of [¹⁹ F]FP2OP

将1mL 1mol/L四正丁基氟化铵的THF溶液加至25mL茄形瓶中，110℃氮气吹干。加入1mL无水乙腈蒸发至干，重复三次。将251mg OTs-P2OP溶于3mL无水乙腈，加至反应瓶中，回流40min。反应液经旋蒸除去溶剂后，用15mL二氯甲烷溶解，并转移至100mL分液漏斗，用20mL水洗涤。有机相用MgSO₄干燥，抽滤后旋蒸滤液，经200-300目的硅胶柱纯化，展开剂为二氯甲烷∶甲醇＝100∶1。最后得到[¹⁹F]FP2OP，为棕色固体。核磁谱图：(¹HNMR，CDCl₃)δ：1.559(s，9H，N(CH₃)₃)，3.759(dt，2H，CH₂CH₂OF)，3.835(t，2H，phenyl-O-CH₂CH₂)，4.092(t，2H，phenyl-OCH₂CH₂)，4.534(dt，2H，CH₂CH₂OF)，5.179(s，2H，CH₂-O-C＝C-Cl)，6.883(d，2H，O-phenyl)，7.262(d，2H，CO-phenyl)，7.659(s，1H，N＝C-H)；(¹⁹FNMR，CDCl₃)δ：-222.86。红外谱图：(IR)/cm^-1：vC＝O：1641，v＝C-O-C：1242，vC-O-C：1091。质谱：C19H24ClFN2O4：计算值：398.1；实际值：399.1。Add 1mL of 1mol/L THF solution of tetra-n-butylammonium fluoride into a 25mL eggplant-shaped bottle, and dry it with nitrogen at 110°C. Add 1 mL of anhydrous acetonitrile and evaporate to dryness, repeating three times. Dissolve 251mg of OTs-P2OP in 3mL of anhydrous acetonitrile, add to the reaction flask, and reflux for 40min. After removing the solvent by rotary evaporation, the reaction solution was dissolved in 15 mL of dichloromethane, transferred to a 100 mL separatory funnel, and washed with 20 mL of water. The organic phase was dried with MgSO₄ . After suction filtration, the filtrate was evaporated and purified through a 200-300 mesh silica gel column. The developing solvent was dichloromethane:methanol=100:1. [¹⁹ F]FP2OP was finally obtained as a brown solid. NMR spectrum: (¹ HNMR, CDCl₃ ) δ: 1.559(s, 9H, N(CH₃ )₃ ), 3.759(dt, 2H, CH₂ CH₂ OF), 3.835(t, 2H, phenyl-O- CH₂ CH₂ ), 4.092 (t, 2H, phenyl-OCH₂ CH₂ ), 4.534 (dt, 2H, CH₂ CH₂ OF), 5.179 (s, 2H, CH₂ -OC=C-Cl), 6.883 (d, 2H, O-phenyl), 7.262 (d, 2H, CO-phenyl), 7.659 (s, 1H, N=CH); (¹⁹ FNMR, CDCl₃ ) δ: -222.86. Infrared spectrum: (IR)/cm^-1 : vC=O: 1641, v=COC: 1242, vC-OC: 1091. Mass Spectrum: C19H24ClFN2O4: Calculated: 398.1; Actual: 399.1.

(3)氟-18标记的哒嗪酮类化合物([¹⁸F]FP2OP)的合成(3) Synthesis of Fluorine-18 Labeled Pyridazinones ([¹⁸ F]FP2OP)

将1.5mL含有6mg K₂CO₃和11mg K_2.2.2的[¹⁸F]F^-溶液加至10mL反应瓶中，120℃氮气吹干，加入0.5mL无水乙腈蒸发至干，重复三次。将2mg OTs-P2OP溶于1mL无水乙腈，加至反应瓶中，90℃插针头反应30分钟。反应结束后冷却至室温，用水稀释至10mL，充分混合后用10mL注射器加入Sep-Pak C₁₈Plus柱，用10mL水淋洗。氮气吹干柱子后，用3mL二氯甲烷淋洗至另一反应瓶中，测定产物放射性活度。Add 1.5 mL of [¹⁸ F]^F -solution containing 6 mg K₂ CO₃ and 11 mg K_2.2.2 into a 10 mL reaction flask, blow dry with nitrogen at 120°C, add 0.5 mL of anhydrous acetonitrile and evaporate to dryness, repeat three times. Dissolve 2mg of OTs-P2OP in 1mL of anhydrous acetonitrile, add to the reaction bottle, and react at 90°C for 30 minutes. After the reaction, cool to room temperature, dilute to 10mL with water, mix well, add to Sep-Pak C₁₈ Plus column with 10mL syringe, rinse with 10mL water. After drying the column with nitrogen, wash it with 3mL of dichloromethane into another reaction bottle, and measure the radioactivity of the product.

60℃氮气吹干上述反应瓶中的二氯甲烷，用0.5mL乙腈溶解后注入C-18反相半制备柱(10×250mm，Venusil MP-C18，Agela Technologies Inc.)。收集保留时间为21.5～22.5min的组分，即为[¹⁸F]FP2OP。HPLC条件为：A相为水，B相为乙腈；淋洗梯度为：0～5min：95％A，5.01～8min：95％～60％A，8.01～25min：60％～10％A，25.01～40min：10％A；流速5mL/min。The dichloromethane in the above reaction flask was blown dry with nitrogen at 60°C, dissolved with 0.5 mL of acetonitrile and injected into a C-18 reversed-phase semi-preparative column (10×250 mm, Venusil MP-C18, Agela Technologies Inc.). The fraction with a retention time of 21.5-22.5 min was collected, which was [¹⁸ F]FP2OP. The HPLC conditions are: phase A is water, phase B is acetonitrile; the eluting gradient is: 0～5min: 95%A, 5.01～8min: 95%～60%A, 8.01～25min: 60%～10%A, 25.01 ~40min: 10%A; flow rate 5mL/min.

通过RP-HPLC鉴定，[¹⁸F]FP2OP的保留时间为22.0min，放射化学纯度大于95％。稳定参考化合物[¹⁹F]FP2OP在相同条件下的保留时间为21.9min。As identified by RP-HPLC, the retention time of [¹⁸ F]FP2OP was 22.0 min, and the radiochemical purity was greater than 95%. The retention time of the stable reference compound [¹⁹ F]FP2OP under the same conditions was 21.9 min.

Claims (2)

1. the pyridazinone compound of class fluoro-18 marks, its general structure is as follows:

Wherein, n=2.

2. the preparation method of the pyridazinone compound of 18 marks of the fluoro-shown in the preparation formula 10, its preparation process is as follows:

A. compound 7 is synthetic:

A. under the ice bath, with an amount of MnO₂Add in an amount of concentrated hydrochloric acid with furfural in batches, 20～100 ℃ of reaction 1～2h, cool overnight, suction filtration, recrystallization obtain compound 9;

B. under the ice bath, with an amount of compound 9, Na₂CO₃Add in the suitable quantity of water with tertiary butyl hydrazonium salt hydrochlorate, stir 1～3h, suction filtration; Precipitation is added in an amount of benzene and the Glacial acetic acid, and 10～60 ℃ of reaction 3～6h obtain compound 8;

C. with an amount of 4-methyl hydroxybenzoate, K₂CO₃, KI and compound 1 be suspended in an amount of pimelinketone, the 12～24h that refluxes obtains compound 2;

D. an amount of compound 2, imidazoles, dimethyl tertiary butyl chloride silane are added among an amount of DMF, 50～100 ℃ of reaction 1～5h obtain compound 3;

E. under the ice bath, an amount of compound 3, lithium aluminium hydride are added in an amount of anhydrous diethyl ether, 0～50 ℃ is stirred 2～3h, obtains compound 4;

F. with an amount of compound 8, Cs₂CO₃, compound 4 adds in an amount of dry DMF, 70～100 ℃ of reaction 12～24h obtain compound 5;

G. tetrahydrofuran solution and the compound 5 with an amount of 1mol/L tetra-n-butyl Neutral ammonium fluoride adds in an amount of tetrahydrofuran (THF), and 10～70 ℃ of reaction 1～2h obtain compound 6;

H. an amount of p-methyl benzene sulfonic chloride, 4-Dimethylamino pyridine, diisopropylethylamine, compound 6 are added to anhydrous CH₂Cl₂In, 5～60 ℃ of reaction 2～4h obtain compound 7;

Its synthetic route is:

B. compound 10 is synthetic

With K₂₂₂, K₂CO₃, [¹⁸F] F^-Add in an amount of anhydrous acetonitrile with compound 7,70～100 ℃ of reaction 20～60min obtain compound 10;

Its synthetic route is:

Related subscript n=2 in the described developed by molecule formula of above steps.