WO2023104009A1 - 一类线粒体靶向的正电子发射或荧光探针、其制备方法及应用 - Google Patents

一类线粒体靶向的正电子发射或荧光探针、其制备方法及应用 Download PDF

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WO2023104009A1
WO2023104009A1 PCT/CN2022/136793 CN2022136793W WO2023104009A1 WO 2023104009 A1 WO2023104009 A1 WO 2023104009A1 CN 2022136793 W CN2022136793 W CN 2022136793W WO 2023104009 A1 WO2023104009 A1 WO 2023104009A1
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程震
陈浩
汪志明
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中国科学院上海药物研究所
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Definitions

  • the invention relates to the technical field of radiopharmaceutical chemistry and fluorescent molecular imaging, in particular to a kind of mitochondria-targeted positron emission or fluorescent probe, its preparation method and application.
  • Mitochondria are the energy supply factories for cell life activities and the places where various physiological and biochemical processes occur.
  • the number of mitochondria is related to the energy demand of cells.
  • a large number of studies have shown that abnormal mitochondrial function is related to various diseases, such as tumors, heart failure, diabetes and so on. Therefore, mitochondrial-targeted imaging can provide a detection basis for the study of mitochondrial function and the diagnosis and treatment of related diseases.
  • DLCs delocalized lipophilic cationic compounds
  • TPP triphenylphosphonium salts
  • 18 F-FBnTP Nature 2019, 575(7782) , 380-384.
  • 18 F-FPEGGBnTP J Labeled Comp Radiopharm 2016,59(3), 117-123
  • 18 F-FTPMP Eur.J.Med.Chem.2016,118,90-97
  • 18 F-FMBTP Mol.Pharm.2014, 11 (11), 3823-3831
  • 18 F-FPTP ACS Med. Chem. Lett.
  • F16 compounds are highly hydrophobic and can cross the hydrophobic barrier of cell membranes and mitochondrial membranes, and are highly enriched in cardiomyocytes and tumor cells with high mitochondrial membrane potential.
  • F16 compounds have fluorescent properties, which are very suitable for low-cost fluorescence Imaging screening, and surgical navigation with clinical fluoroscopic imaging.
  • mitochondria-targeted positron probes of indolevinyl quinolate F16 compounds is of great significance for broadening the field of molecular probes for myocardial perfusion and tumor PET (positron emission computed tomography).
  • One object of the present invention is to provide a class of 18 F or 19 F labeled or substituted indoethylene quinolate salt F16 compounds.
  • Another object of the present invention is to provide a process for the preparation of said compound.
  • Yet another object of the present invention is to provide the use of said compound as a mitochondria-targeted positron emission or fluorescent probe.
  • the present invention adopts the following technical solutions:
  • n 0, 1, 2, 3 or 4;
  • X - is an anion in any form, preferably I - , Br - , BF 4 - or ClO 4 - ;
  • R 1 and R 2 are each independently selected from hydrogen, halogen, cyano, nitro, C1-C6 alkoxy, -NR 4 R 5 , wherein R 4 and R 5 are each independently selected from H, C1-C6 Alkyl, or R 4 and R 5 together with the connected N constitute a substituted or unsubstituted 5-7 membered saturated heterocyclic group, the substituted substituents are selected from C1-C6 alkyl, C1-C6 alkoxy,
  • the 5-7 membered saturated heterocyclic group contains 1 or 2 heteroatoms selected from N, O, S, in particular, -NR 4 R 5 is selected from di(C1-C6 alkyl)amino, pyrrolyl, piperyl Pyridyl, unsubstituted or C1-C6 alkyl substituted piperazinyl, morpholinyl,
  • R 1 and R 2 are each independently:
  • R 3 is 18 F or 19 F.
  • the compound of formula I according to the invention is preferably selected from the following compounds:
  • the present invention further provides the preparation method of described compound of formula I, and its preparation route is as follows:
  • the method is carried out by the following method one or method two,
  • n, R 1 , R 2 , X - are each as defined above, and Hal represents halogen, especially chlorine or bromine.
  • the method includes the steps of:
  • n, R 1 , R 2 , X - are respectively as above, and Hal represents halogen.
  • the present invention provides the preparation steps of 19 F-F16 as follows:
  • the present invention provides the preparation steps of the positron probe 18 F-F16 as follows:
  • the 1,4-lutidine salt may be selected from 1,4-lutidine bromide, 1,4-lutidine iodide, 1,4-lutidine perchlorate , 1,4-lutidine tetrafluoroborate.
  • the present invention also provides the application of the compound of formula I as a mitochondria-targeted positron emission or fluorescent probe, especially in myocardial perfusion and tumor imaging, especially in the preparation of Myocardial perfusion PET imaging agent or tumor PET imaging agent or application in preparation of myocardial perfusion fluorescent imaging agent or tumor fluorescent imaging agent.
  • the 18 F-labeled F16 compound according to the present invention is simple to prepare, has good chemical stability and high radiochemical purity, and can be used as a positron emission probe in the fields of myocardial perfusion and tumor PET imaging.
  • the 19 F-substituted F16 compound according to the present invention has the advantages of simple preparation, good chemical stability, and fluorescence, and can be used as a fluorescent probe in in vivo imaging, for example, in the fields of myocardial perfusion and tumor fluorescence imaging.
  • PET/CT imaging results show that the uptake in the target tissue heart or tumor is relatively high, and it is expected It has been applied in clinic and developed into a new type of myocardial perfusion and tumor PET imaging agent.
  • Figure 1 schematically shows a schematic diagram of imaging of mitochondrial targeting of compounds according to the present invention as positron emission or fluorescent probes in mice;
  • Fig. 2 is the radiochemical purity HPLC spectrum of 18 F-5BEE-F16 prepared in Example 12;
  • Fig. 3 is the radiochemical purity HPLC spectrum of 18 F-5MEF-F16 prepared in Example 13;
  • Fig. 4 is the radiochemical purity HPLC spectrum of 18 F-6FEF-F16 prepared in Example 14;
  • Figure 5 is the HPLC profile of the in vitro stability test of 18 F-5MEF-F16 prepared in Example 13 (a: incubation in PBS for 1 h, b: incubation in PBS for 2 h, c: incubation in FBS for 1 h, d: incubation in FBS for 2 h, );
  • Fig. 6 is micro-PET/CT imaging effect diagram of myocardial perfusion of 18 F-5MEF-F16 prepared in Example 13 in normal Balb/c mice (the arrow indicates the heart);
  • Fig. 7 is a micro-PET/CT imaging effect diagram of 18 F-6FEF-F16 prepared in Example 14 in vivo in 4T1 tumor-bearing mice (arrows indicate tumors);
  • Fig. 8 is a fluorescence imaging diagram of 11 kinds of 19 F-F16 compounds prepared in Examples 1-11 in normal mouse isolated tissues;
  • Fig. 9 is an in vivo fluorescence imaging image of 19 F-EE-F16 prepared in Example 7 in 4T1 tumor-bearing mice (arrows indicate tumors).
  • the raw materials, reagents, and experimental animals used in this application are conventional raw materials, reagents, and experimental animals in this field and are commercially available.
  • the animal experiments comply with the ethical requirements of experimental animals, and the equipment and methods used are all Conventional equipment and methods in the art.
  • Dissolve intermediate 2 (1.56g, 5mmol) of the previous step, triethylamine (0.76g, 7.5mmol) and a small amount of 4-dimethylaminopyridine (61mg, 0.5mmol) in 50mL of dry dichloromethane, under ice-water bath conditions
  • Add p-toluenesulfonyl chloride (1.24g, 6.5mmol) in batches, return to room temperature and react for 4h after the addition is complete, dilute with sodium bicarbonate solution after the reaction, and extract with ethyl acetate for 3 times, the organic layer is washed with dilute hydrochloric acid and saturated brine respectively Washed, dried over anhydrous sodium sulfate, filtered and concentrated, purified by column chromatography to obtain 1.72 g of the labeled precursor compound 3 as a white solid with a yield of 74%.
  • the synthesis method is the same as in Example 1, except that 5-methoxyindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, and 0.22 g of a red solid product is obtained by recrystallization.
  • the synthesis method is the same as in Example 1, except that 5-methoxyindole-3-formaldehyde is used instead of 5-bromoindole-3-formaldehyde, 2-bromoethanol is used instead of 2-chloroethoxyethanol, and orange Yellow solid product 0.22g.
  • the synthesis method is the same as in Example 1, except that 5-fluoroindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, and 1,4-lutidine bromide is used instead of 1,4-dimethyl Pyridine iodide was recrystallized to obtain 0.26 g of an orange solid product.
  • the synthesis method is the same as in Example 1, except that 5-fluoroindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, 2-bromoethanol is used instead of 2-chloroethoxyethanol, and 1,4-di
  • the picoline perchlorate was substituted for 1,4-lutidine iodide, and recrystallized to obtain 0.20 g of an orange solid product.
  • the synthesis method is the same as in Example 1, except that indole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, 2-bromoethanol is used instead of 2-chloroethoxyethanol, and 1,4-lutidine Tetrafluoroborate was substituted for 1,4-lutidine iodide, and recrystallized to obtain 0.26 g of a brown solid product.
  • the synthesis method is the same as in Example 1, except that 5-dimethylaminoindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, and 1,4-lutidine perchlorate is used instead of 1,4 - Lutidine iodide, recrystallized to obtain 0.19 g of a red solid product.
  • the synthesis method is the same as in Example 1, except that 6-fluoroindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, 2-bromoethanol is used instead of 2-chloroethoxyethanol, and 1,4-di
  • the picoline tetrafluoroborate was substituted for 1,4-lutidine iodide, and recrystallized to obtain 0.23 g of a brown solid product.
  • the synthesis method is the same as in Example 1, except that 5-cyanindole-3-carbaldehyde is used instead of 5-bromoindole-3-carbaldehyde, 2-bromoethanol is used instead of 2-chloroethoxyethanol, and orange yellow is obtained by recrystallization Solid product 0.19 g.
  • the QMA column is sequentially passed through 10mL 1M sodium bicarbonate aqueous solution and 10mL deionized water and blown dry.
  • the reaction tube was heated and azeotropically dried by high-purity nitrogen gas at 110°C, then 2 mg of 1,4-lutidine iodide, 2 ⁇ L of piperidine and 0.5 mL of anhydrous acetonitrile were added, and the reaction was sealed at 100°C for 10 minutes. After cooling, add 1.5mL mobile phase to dilute, filter through a 0.22 ⁇ m filter membrane and then separate and purify by HPLC.
  • the effluent of the target product with a retention time of 15.8 min was collected, dried by nitrogen blowing, and diluted with 1 mL of sterile normal saline for later use.
  • the decay-corrected specific activity of 18 F-5MEF-F16 is 127.4 ⁇ 25.0 GBq/ ⁇ mol, and the radiochemical purity is >98% as shown in Figure 3 .
  • the radiolabeled synthesis of 18 F-6FEF-F16 is the same as in Example 12, except that 1,4-lutidine tetrafluoroborate is used, the labeled precursor compound 3 prepared in Example 10 is used, and the dosage is .
  • the reaction conditions were consistent with the HPLC separation conditions.
  • the effluent of the target product with a retention time of 11.5 min was collected, dried by nitrogen blowing, and diluted with 1 mL of sterile normal saline for use.
  • the decay-corrected specific activity of 18 F-6FEF-F16 is 93.8 ⁇ 12.6 GBq/ ⁇ mol, and the radiochemical purity is >98% as shown in Figure 4 .
  • Table 1 Table of optical properties of 19 F-F16 compounds
  • Test Example 2 In vitro stability test of 18 F-5MEF-F16
  • Test Example 3 Micro-PET/CT Imaging Application of 18 F-5MEF-F16 in Myocardial Perfusion in Mice
  • mice Take 3 normal commercially available SPF Balb/c mice and inject 100-150 ⁇ Ci dose of 18 F-5MEF-F16 probe prepared in Example 13 through the tail vein respectively, and maintain with 2.0% isoflurane-oxygen mixed gas
  • the mice were under anesthesia, and PET/CT scanning (Siemens Inveon microPET) was performed synchronously at the beginning of the injection, dynamic imaging images were collected within 120 minutes, ROIs were drawn, and %ID/g was calculated. The results are shown in Figure 6 and Table 2.
  • Test Example 4 Micro-PET/CT imaging application of 18 F-6FEF-F16 in tumor-bearing mice
  • PET/CT imaging of 18 F-6FEF-F16 in tumor-bearing mice showed that the probe molecules were obviously concentrated in the tumor site over time, and reached the highest uptake value of 16.02 ⁇ 3.91% ID in 1 hour /g, significantly higher than that of muscle, heart, lung and other organs and tissues, the tumor/muscle ratio was 7.49.
  • PET/CT imaging shows that 18 F-6FEF-F16 can specifically target the tumor site, has a good target/non-target ratio, and the tumor focus is clearly visible, which has a good research and application prospect.
  • Test Example 5 Application of 19 F-F16 in Fluorescence Imaging of Mouse Isolated Tissues
  • some 19 F-F16 compounds are obviously distributed in heart tissue, and have a higher target/non-target ratio, indicating that the compounds of the present invention can be used as probes for myocardial perfusion imaging.
  • Test Example 6 Application of 19 F-EE-F16 in live imaging of tumor-bearing mice

Abstract

本发明涉及一类线粒体靶向的正电子发射或荧光探针、其制备方法及应用。该探针结构如式(I)所示，为18F或19F取代的吲哚乙烯喹啉盐类(F16)化合物。该类化合物具有制备简单、化学稳定性好、放射化学纯度高，具有荧光等优点，可作为线粒体成像相关的心肌灌注和肿瘤PET显像剂或荧光显像剂应用在放射性药物化学和荧光分子影像技术领域中。

Description

一类线粒体靶向的正电子发射或荧光探针、其制备方法及应用 技术领域
本发明涉及放射性药物化学和荧光分子影像技术领域，具体地说，涉及一类线粒体靶向的正电子发射或荧光探针、其制备方法及应用。
背景技术
线粒体是细胞生命活动的能量供应工厂，是多种生理生化过程的发生场所。线粒体的数量与细胞的能量需求有关，新陈代谢旺盛的器官和组织，如心脏、肿瘤组织，其细胞内含有大量线粒体。大量研究表明，线粒体功能异常与多种疾病相关，如肿瘤、心衰、糖尿病等。因此，线粒体靶向成像可以为研究线粒体功能和相关疾病的诊疗提供检测依据。
心脏和肿瘤组织代谢活跃，为其供应能量的线粒体具有更高的线粒体膜电位，一些离域亲脂性阳离子化合物(DLCs)在线粒体膜电位的驱使下，利用分子亲脂性特性进入线粒体基质内，达到线粒体富集效果。目前以线粒体为靶点的亲脂性阳离子在肿瘤分子影像探针、抗肿瘤药物和心肌灌注成像研究领域，已成为一大热点。但上述亲脂性阳离子探针多以荧光试剂居多，正电子探针开发相对较少，且多以三苯基膦鎓盐(TPP)为主，如 18F-FBnTP(Nature 2019,575(7782),380-384.)、 18F-FPEGBnTP(J Labelled Comp Radiopharm 2016,59(3),117-123)、 18F-FTPMP(Eur.J.Med.Chem.2016,118,90-97)、 18F-FMBTP(Mol.Pharm.2014,11(11),3823-3831)和 18F-FPTP(ACS Med.Chem.Lett.2014,5(10),1124-1128)等，其它结构如罗丹明衍生物为代表的 18F-FERhB(Nucl.Med.Biol.2010,37(3),365-370)、 18F-Rhodamin 6G(Medchemcomm 2017,8(10),1891-1896)亦有报道。
但广泛应用于肿瘤线粒体靶向的吲哚乙烯喹啉盐F16类化合物(Cancer Cell 2002,2(1),29-42；Chem.Commun.(Camb.)2014,50(64),8919-8922；Chemical Science 2019,10(34),7946-7951)却未有正电子探针研究报道。F16类化合物具有高度疏水性，可以跨过细胞膜和线粒体膜的疏水屏障，在线粒体膜电位较高的心肌细胞和肿瘤细胞高度富集，同时F16类化合物本身具备荧光特性，非常适合低成本的荧光成像筛选，以及临床荧光成像手术导航。因此，开发线粒体靶向的吲哚乙烯喹啉盐F16类化合物的正电子探针为拓宽心肌灌注和肿瘤PET(正电子发射型计算机断层显像)分子探针领域具有重要意义。
发明内容
本发明的一个目的是提供一类 18F或 19F标记或取代的吲哚乙烯喹啉盐F16类化合物。
本发明的另一个目是提供所述化合物的制备方法。
本发明的再一个目的是提供所述化合物作为线粒体靶向的正电子发射或荧光探针的应用。
为实现上述目的，本发明采用以下技术方案：
根据本发明的一个方面，提供一种式I化合物：
Figure PCTCN2022136793-appb-000001
其中：n为0，1，2，3或4；
X -为任意形式阴离子，优选为I -，Br -，BF 4 -或ClO 4 -；
R 1和R 2各自独立地选自氢、卤素、氰基、硝基、C1-C6烷氧基、-NR 4R 5，其中，R 4和R 5各自独立地选自H、C1-C6烷基，或者R 4和R 5与相连的N共同构成取代或未取代的5-7元饱和杂环基，所述取代的取代基选自C1-C6烷基、C1-C6烷氧基，所述5-7元饱和杂环基含有1或2个选自N、O、S的杂原子，特别地，-NR 4R 5选自二(C1-C6烷基)氨基、吡咯基、哌啶基、未取代或C1-C6烷基取代的哌嗪基、吗啉基，
优选R 1和R 2各自独立地为：
Figure PCTCN2022136793-appb-000002
R 3为 18F或 19F。
具体地，根据本发明的式I化合物优选选自以下化合物：
Figure PCTCN2022136793-appb-000003
Figure PCTCN2022136793-appb-000004
本发明进一步提供所述的式I化合物的制备方法，其制备路线如下：
Figure PCTCN2022136793-appb-000005
所述方法通过如下方法一或方法二进行，
方法一
包括如下步骤：
(a)吲哚3-甲醛化合物1和Hal-(CH 2CH 2O) nC 2H 5OH发生亲核取代反应得到化合物2；
(b)化合物2与对甲苯磺酰氯发生酯化反应得到化合物3；
(c)化合物3在氨基聚醚(K2.2.2)存在下与 18F-F -发生亲核取代反应得到 18F取代的化合物 18F-4；
(d)化合物 18F-4与1,4-二甲基吡啶盐发生Knoevenagel缩合反应得到终产物 18F-F16；
方法二
包括如下步骤：
(a)吲哚3-甲醛化合物1和Hal-(CH 2CH 2O) nC 2H 5OH发生亲核取代反应得到化合物2；
(b)化合物2与对甲苯磺酰氯发生酯化反应得到化合物3；
(e)化合物3与四丁基氟化铵发生亲核取代反应得到 19F取代的化合物 19F-4，
(f)化合物 19F-4与1,4-二甲基吡啶盐发生Knoevenagel缩合反应得到终产物 19F-F16；
其中n，R 1，R 2，X -分别如上所定义，Hal表示卤素，特别是氯或溴。
在一实施方式中，所述方法包括如下步骤：
(1)吲哚3-甲醛化合物1和碳酸钾加入到N,N-二甲基甲酰胺中，并加入Hal-(CH 2CH 2O) nC 2H 5OH(例如2-溴乙醇，2-氯乙氧基乙醇，2-[(2-氯乙氧基)乙氧基]乙 醇，或2-[2-[2-(2-氯乙氧基)乙氧基]乙氧基]乙醇)和少量碘化钾，90℃搅拌反应8h，反应完毕后冷却、稀释、对稀释溶液萃取、洗涤有机层、干燥、过滤、浓缩并纯化，得到中间体化合物2；
(2)将化合物2、三乙胺和少量4-二甲氨基吡啶溶于干燥二氯甲烷中，冰水浴条件下分批次加入对甲苯磺酰氯，加入完毕后恢复室温反应4h，反应完毕后稀释、对稀释溶液萃取、洗涤有机层、干燥，过滤并浓缩纯化，得到标记前体化合物3；
(3-1)将化合物3和四丁基氟化铵在无水四氢呋喃或无水乙腈溶液中加热90℃反应，得 19F取代的化合物 19F-4，或
(3-2)将氨基聚醚(K2.2.2)、碳酸钾、 18F-F -和化合物3在无水乙腈中100℃反应10min，得到 18F取代的化合物 18F-4；
(4-1)将上述(3-1)得到的 19F取代的化合物 19F-4、1倍当量的1,4-二甲基吡啶盐和0.2当量的哌啶在氮气保护下无水甲醇中回流加热过夜，冷却后重结晶得到终产物 19F-F16，或
(4-2)将上述(4-1)得到的 18F取代的化合物 18F-4、1,4-二甲基吡啶盐和哌啶在无水乙腈中100℃反应10min，冷却后经分离纯化得到终产物 18F-F16，
其中n，R 1，R 2，X -分别如上所述，Hal表示卤素。
在一实施方式中，本发明提供所述的 19F-F16的制备步骤如下：
(1)将吲哚3-甲醛化合物1和碳酸钾加入到20mL N,N-二甲基甲酰胺中，并加入2-溴乙醇或2-氯乙氧基乙醇或2-[(2-氯乙氧基)乙氧基]乙醇或2-[2-[2-(2-氯乙氧基)乙氧基]乙氧基]乙醇和少量碘化钾，90℃搅拌反应8h，反应完毕待冷却后，用饱和氯化铵溶液稀释反应液，并用乙酸乙酯萃取3次，有机层用饱和盐水洗涤，无水硫酸钠干燥，过滤并浓缩经过柱层析纯化，得到中间体化合物2；
(2)将化合物2、三乙胺和少量4-二甲氨基吡啶溶于50mL干燥二氯甲烷中，冰水浴条件下分批次加入对甲苯磺酰氯，加入完毕后恢室温反应4h，反应完毕后碳酸氢钠溶液稀释，并用乙酸乙酯萃取3次，有机层分别用稀盐酸和饱和盐水洗涤，无水硫酸钠干燥，过滤并浓缩经过柱层析纯化，得到标记前体化合物3；
(3)将化合物3和四丁基氟化铵在无水四氢呋喃或无水乙腈溶液中加热反应，可得 19F取代的吲哚-3-甲醛化合物 19F-4；
(4)将上述得到的 19F取代化合物 19F-4、1,4-二甲基吡啶盐和0.2当量的哌啶在氮气保护下无水甲醇中回流加热过夜，冷却后重结晶得到终产物 19F-F16。
在另一实施方式中，本发明提供所述的正电子探针 18F-F16的制备步骤如下：
(1)将吲哚3-甲醛化合物1和碳酸钾加入到20mL N,N-二甲基甲酰胺中，并加入 2-溴乙醇或2-氯乙氧基乙醇或2-[(2-氯乙氧基)乙氧基]乙醇或2-[2-[2-(2-氯乙氧基)乙氧基]乙氧基]乙醇和少量碘化钾，90℃搅拌反应8h，反应完毕待冷却后，用饱和氯化铵溶液稀释反应液，并用乙酸乙酯萃取3次，有机层用饱和盐水洗涤，无水硫酸钠干燥，过滤并浓缩经过柱层析纯化，得到中间体化合物2；
(2)将化合物2、三乙胺和少量4-二甲氨基吡啶溶于50mL干燥二氯甲烷中，冰水浴条件下分批次加入对甲苯磺酰氯，加入完毕后恢室温反应4h，反应完毕后碳酸氢钠溶液稀释，并用乙酸乙酯萃取3次，有机层分别用稀盐酸和饱和盐水洗涤，无水硫酸钠干燥，过滤并浓缩经过柱层析纯化，得到标记前体化合物3；
(3’)将氨基聚醚(K2.2.2)、碳酸钾、 18F-F -和前体化合物3在无水乙腈中100℃反应10min，得到 18F标记化合物4；
(4’)将上述得到的 18F标记化合物4、1,4-二甲基吡啶盐和哌啶在无水乙腈中100℃反应10min，冷却后经半制备高效液相分离得到标记终产物 18F-F16，其可再经氮吹干燥，加入1mL无菌生理盐水稀释备用。
根据本发明，1,4-二甲基吡啶盐可以选自1,4-二甲基吡啶溴化物、1,4-二甲基吡啶碘化物、1,4-二甲基吡啶高氯酸盐、1,4-二甲基吡啶四氟硼酸盐。
根据本发明的另一方面，本发明还提供所述的式I化合物作为线粒体靶向的正电子发射或荧光探针的应用，特别是在心肌灌注和肿瘤显像中的应用，尤其是在制备心肌灌注PET显像剂或肿瘤PET显像剂或者在制备心肌灌注荧光显像剂或肿瘤荧光显像剂中的应用。
根据本发明的 18F标记的F16类化合物，制备简单、化学稳定性好、放射化学纯度高、能够作为正电子发射探针应用在心肌灌注和肿瘤PET显像领域。
根据本发明的 19F取代的F16类化合物，制备简单、化学稳定性好、具有荧光等优点，能够作为荧光探针应用在活体成像中，例如在心肌灌注和肿瘤荧光显像领域。
该类化合物具有制备简单、化学稳定性好、放射化学纯度高，具有荧光等优点，在 18F标记后，PET/CT显像结果中表明，在靶组织心脏或肿瘤中摄取较高，其有望应用于临床，发展为一类新型的心肌灌注和肿瘤PET显像剂。
附图说明
图1示意性地示出了根据本发明的化合物作为正电子发射或荧光探针在小鼠中的线粒体靶向的显像示意图；
图2为实施例12制备的 18F-5BEE-F16的放化纯度HPLC图谱；
图3为实施例13制备的 18F-5MEF-F16的放化纯度HPLC图谱；
图4为实施例14制备的 18F-6FEF-F16的放化纯度HPLC图谱；
图5为实施例13制备的 18F-5MEF-F16的体外稳定性实验HPLC图谱(a：PBS中 孵育1h，b：PBS中孵育2h，c：FBS中孵育1h，d：FBS中孵育2h，)；
图6为实施例13制备的 18F-5MEF-F16在正常Balb/c小鼠体内心肌灌注micro-PET/CT显像效果图(箭头指示为心脏)；
图7为实施例14制备的 18F-6FEF-F16在4T1荷瘤小鼠体内肿瘤micro-PET/CT显像效果图(箭头指示为肿瘤)；
图8为实施例1-11所制备的11种 19F-F16化合物在正常小鼠离体组织荧光成像图；
图9为实施例7制备的 19F-EE-F16在4T1荷瘤小鼠活体荧光成像图(箭头指示为肿瘤)。
具体实施方式
除特殊说明外，本申请中所采用的原料、试剂、试验动物等均为本领域常规原料、试剂、试验动物且市售可得，动物试验遵守实验动物伦理要求，所采用的设备和方法均为本领域常规设备和方法。
实施例1： 19F-5BEE-F16的制备
(1)合成实施例1中间体2
将5-溴吲哚-3-甲醛(2.24g，10mmol)和碳酸钾(2.07g，15mmol)加入到20mL N,N-二甲基甲酰胺中，并加入2-氯乙氧基乙醇(1.62g，13mmol)和少量碘化钾(83mg，0.5mmol)，90℃搅拌反应8h，反应完毕待冷却后，用饱和氯化铵溶液稀释反应液，并用乙酸乙酯萃取3次，有机层用饱和盐水洗涤，无水硫酸钠干燥，过滤并浓缩经过柱层析纯化，得到白色固体2.5g，产率80％。 1H NMR(400MHz,CDCl 3)δ9.93(s,1H),8.45(d,J＝1.9Hz,1H),7.81(s,1H),7.41(dd,J＝8.7,2.0Hz,1H),7.25(d,J＝8.7Hz,1H),4.34(t,J＝5.1Hz,2H),3.87(t,J＝5.1Hz,2H),3.72–3.65(m,2H),3.53(t,J＝4.7Hz,2H),1.90(t,J＝5.8Hz,1H). 13C NMR(101MHz,CDCl 3)δ184.4,139.7,136.0,127.0,126.8,124.8,117.7,116.6,111.4,72.6,69.3,61.7,47.3.结构如下：
Figure PCTCN2022136793-appb-000006
(2)合成实施例1中间体3
将上步中间体2(1.56g，5mmol)、三乙胺(0.76g，7.5mmol)和少量4-二甲氨基吡啶(61mg，0.5mmol)溶于50mL干燥二氯甲烷中，冰水浴条件下分批次加入对甲苯磺酰氯(1.24g，6.5mmol)，加入完毕后恢复室温反应4h，反应完毕后碳酸氢钠溶液稀释，并用乙酸乙酯萃取3次，有机层分别用稀盐酸和饱和盐水洗涤，无水硫酸钠 干燥，过滤并浓缩经过柱层析纯化，得到标记前体化合物3为白色固体1.72g，产率74％。 1H NMR(400MHz,CDCl 3)δ9.93(s,1H),8.46(d,J＝2.0Hz,1H),7.77(s,1H),7.72(d,J＝8.3Hz,2H),7.38(dd,J＝8.8,2.0Hz,1H),7.31(d,J＝8.1Hz,2H),7.21(d,J＝8.7Hz,1H),4.29(t,J＝5.0Hz,2H),4.11–4.06(m,2H),3.80(t,J＝5.0Hz,2H),3.63–3.57(m,2H),2.44(s,3H). 13C NMR(101MHz,CDCl 3)δ184.6,145.1,140.0,135.9,132.8,129.9,127.8,126.9,126.8,124.9,117.7,116.5,111.4,69.5,68.9,68.8,47.1,21.7.结构如下：
Figure PCTCN2022136793-appb-000007
(3)合成实施例1中间体 19F-4
将上步中间体3(0.93g，2mmol)、四丁基氟化铵(6mmol)溶于10mL无水乙腈中，90℃加热反应4h，反应结束后，减压旋蒸除去溶剂，经过柱层析纯化，得到标记前体化合物 19F-4为淡黄色固体0.54g，产率86％。 1H NMR(400MHz,CDCl 3)δ9.86(s,1H),8.38(d,J＝2.0Hz,1H),7.76(s,1H),7.33(dd,J＝8.7,2.0Hz,1H),7.19(d,J＝8.7Hz,1H),4.51(t,J＝3.7Hz,1H),4.39(t,J＝3.9Hz,1H),4.28(t,J＝5.0Hz,2H),3.83(t,J＝5.0Hz,2H),3.65(t,J＝3.9Hz,1H),3.57(t,J＝3.9Hz,1H). 13C NMR(101MHz,CDCl 3)δ184.5,140.2,136.0,126.8,126.7,124.6,117.5,116.4,111.6,83.8,82.1,70.6,70.4,69.5,47.2.结构如下：
Figure PCTCN2022136793-appb-000008
(4)合成实施例1产物 19F-5BEE-F16
将上述 19F-4化合物(0.31g，1mmol)分别与1倍当量1,4-二甲基吡啶碘化物和0.2倍当量哌啶加入到10mL无水甲醇中，氮气保护下反应液回流加热过夜，通过TLC监控反应。待反应结束后，反应液置于-20℃冰箱冷却，伴有沉淀物生成，过滤收集沉淀物，冷甲醇洗涤，最后经乙腈重结晶得橘黄色固体产物0.25g，产率47％。 1H NMR(400MHz,DMSO-d 6)δ8.72(d,J＝6.6Hz,2H),8.36(d,J＝1.9Hz,1H),8.23(d,J＝16.3Hz,1H),8.17(d,J＝6.8Hz,2H),8.06(s,1H),7.65(d,J＝8.7Hz,1H),7.43(dd,J＝8.7,1.9Hz,1H),7.27(d,J＝16.3Hz,1H),4.57–4.51(m,1H),4.46(t,J＝5.1Hz,2H),4.43–4.39(m,1H),4.19(s,3H),3.82(t,J＝5.1Hz,2H),3.73–3.65(m,1H),3.63–3.58(m,1H). 13C  NMR(101MHz,DMSO-d 6)δ154.3,144.8,136.6,135.7,135.0,127.8,125.8,123.0,122.4,121.6,118.2,114.8,113.8,112.7,84.3,82.6,70.1,70.0,69.5,46.8,46.7.HRMS(ESI)m/z:[M-I] +calculated for C 20H 21BrFN 2O:403.0816,405.0796,found:403.0815,405.0792.结构如下：
Figure PCTCN2022136793-appb-000009
实施例2： 19F-5BEF-F16的制备
合成方法同实施例1，不同在于，用2-溴乙醇代替2-氯乙氧基乙醇，重结晶得橘黄色固体产物0.21g。 1H NMR(400MHz,DMSO-d 6)δ8.73(d,J＝6.6Hz,2H),8.39(d,J＝1.9Hz,1H),8.24(d,J＝16.3Hz,1H),8.18(d,J＝6.6Hz,2H),8.08(s,1H),7.65(d,J＝8.7Hz,1H),7.45(dd,J＝8.8,1.8Hz,1H),7.30(d,J＝16.3Hz,1H),4.84(t,J＝4.6Hz,1H),4.72(t,J＝4.5Hz,1H),4.67(t,J＝4.6Hz,1H),4.60(t,J＝4.6Hz,1H),4.20(s,3H). 13C NMR(101MHz,DMSO-d 6)δ154.3,144.8,136.5,135.3,134.8,127.9,126.0,123.0,122.5,118.6,114.9,113.7,113.0,83.8,82.2,47.3,47.1,46.9.HRMS(ESI)m/z:[M-I] +calculated for C 18H 17BrFN 2:359.0554,361.0534,found:359.0553,361.0526.
实施例3： 19F-5MEE-F16的制备
合成方法同实施例1，不同在于，采用5-甲氧基吲哚-3-甲醛代替5-溴吲哚-3-甲醛，重结晶得红色固体产物0.22g。 1H NMR(400MHz,DMSO-d 6)δ8.67(d,J＝6.7Hz,2H),8.23(d,J＝16.2Hz,1H),8.18–8.09(m,2H),7.96(s,1H),7.61(d,J＝2.5Hz,1H),7.56(d,J＝8.9Hz,1H),7.18(d,J＝16.2Hz,1H),6.95(dd,J＝8.9,2.4Hz,1H),4.58–4.50(m,1H),4.46–4.38(m,3H),4.17(s,3H),3.88(s,3H),3.81(t,J＝5.1Hz,2H),3.70–3.65(m,1H),3.63–3.57(m,1H). 13C NMR(101MHz,DMSO-d 6)δ155.8,154.6,144.5,136.1,135.5,132.9,126.8,122.0,116.8,112.9,112.6,112.4,103.6,84.3,82.6,70.1,70.0,69.5,56.3,46.7.HRMS(ESI)m/z:[M-I] +calculated for C 21H 24FN 2O 2:355.1817,found:355.1812.
实施例4： 19F-5MEE-F16的制备
合成方法同实施例1，不同在于，采用5-甲氧基吲哚-3-甲醛代替5-溴吲哚-3-甲醛，2-溴乙醇代替2-氯乙氧基乙醇，重结晶得橘黄色固体产物0.22g。 1H NMR(400MHz,DMSO-d 6)δ8.68(d,J＝6.9Hz,2H),8.23(d,J＝16.2Hz,1H),8.17–8.09(m,2H),7.99(s,1H),7.62(d,J＝2.4Hz,1H),7.56(d,J＝8.9Hz,1H),7.21(d,J＝16.2Hz,1H),6.97(dd,J＝ 8.9,2.4Hz,1H),4.83(t,J＝4.6Hz,1H),4.71(t,J＝4.6Hz,1H),4.62(t,J＝4.6Hz,1H),4.55(t,J＝4.7Hz,1H),4.18(s,3H),3.88(s,3H). 13C NMR(101MHz,DMSO-d 6)δ155.8,154.5,144.6,135.9,135.0,132.8,126.9,122.1,117.2,113.2,112.8,112.4,103.6,83.8,82.1,56.3,47.2,47.0,46.7.HRMS(ESI)m/z:[M–I] +calculated for C 19H 20FN 2O:311.1555,found:311.1551.
实施例5： 19F-5FEE-F16的制备
合成方法同实施例1，不同在于，采用5-氟吲哚-3-甲醛代替5-溴吲哚-3-甲醛，和1,4-二甲基吡啶溴化物代替1,4-二甲基吡啶碘化物，重结晶得橘黄色固体产物0.26g。 1H NMR(400MHz,DMSO-d 6)δ8.70(d,J＝6.6Hz,2H),8.20(d,J＝16.3Hz,1H),8.14(d,J＝6.9Hz,2H),8.04(s,1H),8.00(dd,J＝10.1,2.5Hz,1H),7.69(dd,J＝9.0,4.5Hz,1H),7.26(d,J＝16.3Hz,1H),7.17(td,J＝9.2,2.5Hz,1H),4.57–4.51(m,1H),4.47(t,J＝5.1Hz,2H),4.43–4.39(m,1H),4.18(s,3H),3.82(t,J＝5.0Hz,2H),3.72–3.65(m,1H),3.64–3.58(m,1H). 13C NMR(121MHz,DMSO-d 6)δ159.9,158.0,154.5,144.7,136.7,135.5,134.6,126.4,126.3,122.3,117.7,113.2,113.1,113.0,113.0,111.5,111.3,106.3,106.1,84.1,82.8,70.1,70.0,69.5,46.8,46.8.HRMS(ESI)m/z:[M-Br] +calculated for C 20H 21F 2N 2O:343.1617,found:343.1614.
实施例6： 19F-5FEF-F16的制备
合成方法同实施例1，不同在于，采用5-氟吲哚-3-甲醛代替5-溴吲哚-3-甲醛，2-溴乙醇代替2-氯乙氧基乙醇，和1,4-二甲基吡啶高氯酸盐代替1,4-二甲基吡啶碘化物，重结晶得橘黄色固体产物0.20g。 1H NMR(400MHz,DMSO-d 6)δ8.71(d,J＝6.9Hz,2H),8.21(d,J＝16.3Hz,1H),8.15(d,J＝7.0Hz,2H),8.07(s,1H),8.02(dd,J＝10.1,2.5Hz,1H),7.69(dd,J＝9.0,4.5Hz,1H),7.29(d,J＝16.4Hz,1H),7.19(td,J＝9.1,2.5Hz,1H),4.84(t,J＝4.5Hz,1H),4.72(t,J＝4.5Hz,1H),4.67(t,J＝4.6Hz,1H),4.60(t,J＝4.6Hz,1H),4.19(s,3H). 13C NMR(126MHz,DMSO-d 6)δ159.9,158.1,154.4,144.8,136.3,135.2,134.5,126.5,126.4,122.4,118.1,113.5,113.5,113.0,112.9,111.6,111.4,106.3,106.1,83.7,82.3,47.3,47.2,46.9.HRMS(ESI)m/z:[M–ClO 4] +calculated for C 18H 17F 2N 2:299.1355,found:299.1353.
实施例7： 19F-EE-F16的制备
合成方法同实施例1，不同在于，采用吲哚-3-甲醛代替5-溴吲哚-3-甲醛，重结晶得棕色固体产物0.26g。 1H NMR(400MHz,DMSO-d 6)δ8.67(d,J＝6.7Hz,2H),8.22(d,J＝16.2Hz,1H),8.18(dd,J＝7.3,1.6Hz,1H),8.13(d,J＝7.0Hz,2H),7.98(s,1H),7.66(dd,J＝7.6,1.4Hz,1H),7.37–7.23(m,3H),4.57–4.52(m,1H),4.46(t,J＝5.1Hz,2H),4.44–4.40(m,1H),4.17(s,3H),3.83(t,J＝5.1Hz,2H),3.71–3.65(m,1H),3.63–3.58 (m,1H). 13C NMR(101MHz,DMSO-d 6)δ154.5,144.7,137.9,136.1,135.7,125.9,123.4,122.1,121.9,121.1,117.4,113.2,111.7,84.3,82.6,70.1,70.0,69.5,46.7,46.4.HRMS(ESI)m/z:[M-I] +calculated for C 20H 22FN 2O:325.1711,found:325.1708.
实施例8： 19F-EF-F16的制备
合成方法同实施例1，不同在于，采用吲哚-3-甲醛代替5-溴吲哚-3-甲醛，2-溴乙醇代替2-氯乙氧基乙醇，和1,4-二甲基吡啶四氟硼酸盐代替1,4-二甲基吡啶碘化物，重结晶得棕色固体产物0.26g。 1H NMR(400MHz,DMSO-d 6)δ8.71(d,J＝6.8Hz,2H),8.24(d,J＝16.3Hz,1H),8.20(dd,J＝7.3,1.5Hz,1H),8.15(d,J＝6.9Hz,2H),8.01(s,1H),7.67(d,J＝7.9Hz,1H),7.39–7.25(m,3H),4.85(t,J＝4.6Hz,1H),4.73(t,J＝4.6Hz,1H),4.68(t,J＝4.7Hz,1H),4.61(t,J＝4.6Hz,1H),4.18(s,3H). 13C NMR(101MHz,DMSO-d 6)δ154.0,144.2,137.4,135.4,134.8,125.5,125.5,123.1,121.8,121.5,120.6,117.3,113.1,111.2,83.1,81.8,46.5,46.4,46.3.HRMS(ESI)m/z:[M–BF 4] +calculated for C 18H 18FN 2:281.1449,found:281.1448.
实施例9： 19F-5NEE-F16的制备
合成方法同实施例1，不同在于，采用5-二甲氨基吲哚-3-甲醛代替5-溴吲哚-3-甲醛，和1,4-二甲基吡啶高氯酸盐代替1,4-二甲基吡啶碘化物，重结晶得红色固体产物0.19g。 1H NMR(400MHz,DMSO-d 6)δ8.68(d,J＝6.7Hz,2H),8.23(d,J＝16.2Hz,1H),8.14(d,J＝6.8Hz,2H),7.98(s,1H),7.65(d,J＝2.5Hz,1H),7.46(d,J＝8.9Hz,1H),7.18(d,J＝16.2Hz,1H),6.99(dd,J＝8.9,2.4Hz,1H),4.58–4.50(m,1H),4.46–4.38(m,3H),4.17(s,3H),3.88(s,3H),3.81(t,J＝5.1Hz,2H),3.70–3.65(m,1H),3.63–3.57(m,1H),3.02(s,6H). 13C NMR(101MHz,DMSO-d 6)δ155.7,153.6,142.5,136.7,135.6,132.2,123.8,122.1,117.8,112.6,112.3,112.5,103.6,84.3,82.6,70.1,70.0,69.5,46.7,40.8.HRMS(ESI)m/z:[M–ClO 4] +calculated for C 22H 27FN 3O:368.2133,found:368.2132.
实施例10： 19F-6FEF-F16的制备
合成方法同实施例1，不同在于，采用6-氟吲哚-3-甲醛代替5-溴吲哚-3-甲醛，2-溴乙醇代替2-氯乙氧基乙醇，和1,4-二甲基吡啶四氟硼酸盐代替1,4-二甲基吡啶碘化物，重结晶得棕色固体产物0.23g。 1H NMR(400MHz,DMSO-d 6)δ8.73(d,J＝6.6Hz,2H),8.25–8.18(m,2H),8.16(d,J＝6.9Hz,2H),8.02(s,1H),7.60(dd,J＝10.2,2.4Hz,1H),7.33(d,J＝16.3Hz,1H),7.16(td,J＝9.2,2.4Hz,1H),4.84(t,J＝4.6Hz,1H),4.72(t,J＝4.5Hz,1H),4.65(t,J＝4.7Hz,1H),4.58(t,J＝4.7Hz,1H),4.20(s,3H).13C NMR(101MHz,DMSO-d 6)δ161.2,158.9,154.3,144.8,138.3,138.2,135.6,135.3,122.6,122.4,122.3,122.2,118.3,113.7,110.2,110.0,98.6,98.3,83.8,82.1,47.2,47.0,46.9.HRMS(ESI)m/z:[M–BF 4] +calculated for C 18H 17F 2N 2:299.1355,found:299.1352.
实施例11： 19F-5CNEF-F16的制备
合成方法同实施例1，不同在于，采用5-氰基吲哚-3-甲醛代替5-溴吲哚-3-甲醛，2-溴乙醇代替2-氯乙氧基乙醇，重结晶得橘黄色固体产物0.19g。 1H NMR(400MHz,DMSO-d 6)δ8.82–8.74(m,3H),8.26(d,J＝16.4Hz,1H),8.22–8.17(m,3H),7.88(d,J＝8.6Hz,1H),7.71(dd,J＝8.6,1.5Hz,1H),7.44(d,J＝16.4Hz,1H),4.86(t,J＝4.6Hz,1H),4.78–4.71(m,2H),4.67(t,J＝4.6Hz,1H),4.22(s,3H). 13C NMR(101MHz,DMSO-d 6)δ154.0,145.0,139.4,136.2,134.0,126.2,126.1,125.7,122.7,120.6,119.7,114.0,113.0,103.9,83.8,82.2,47.2,47.0,47.0.HRMS(ESI)m/z:[M-I] +calculated for C 19H 17FN 3:306.1402,found:306.1400.
实施例12： 18F-5BEE-F16的放化标记
QMA柱依次使用10mL 1M碳酸氢钠水溶液和10mL去离子水通过并吹干，回旋加速器生产 18F-氟离子富集在QMA柱上，用1mL淋洗液(含13mg K2.2.2和3mg碳酸钾，乙腈/水＝9/1)将 18F-氟离子从QMA柱上洗脱至5mL反应管中。吹入高纯氮气110℃加热共沸吹干，再加入1.5mL无水乙腈共沸吹干，重复两次保证反应体系无水。将2mg实施例1中制备的标记前体化合物3加入到反应管中，100℃密闭反应10分钟，反应管冷却后吸出倒入10mL水中，负载于C18Sep-pak柱，后以20mL纯水淋洗杂质，再以1mL无水乙腈洗脱收集于反应管中。反应管经高纯氮气110℃加热共沸吹干，再加入2mg 1,4-二甲基吡啶碘化物、2μL哌啶和0.5mL无水乙腈，100℃密闭反应10分钟。冷却后加入1.5mL流动相稀释，0.22μm滤膜过滤后通过HPLC分离纯化，分离条件：岛津XBridge BEH C18反向柱(10×250mm,5μm)；流动相：A＝PBS水溶液，B＝乙腈；梯度：0-5min，25％B，5-25min，25％B-80％B；流速：3mL/min。收集保留时间11.8min目标产物的流出液，再经氮吹干燥，得到 18F-5BEE-F16。向其中加入1mL无菌生理盐水稀释备用。 18F-5BEE-F16衰变校正后比活度103.2±30.6GBq/μmol，放化纯度如图2所示>98％。
实施例13： 18F-5MEF-F16的放化标记
18F-5MEF-F16的放射性标记合成同实施例12，其投药量、反应条件一致。不同在于，使用实施例4中制备的标记前体化合物3，HPLC分离条件：岛津XBridge BEH C18反向柱(10×250mm,5μm)；流动相：A＝PBS水溶液，B＝乙腈；梯度：0-5min，20％B，5-25min，20％B-75％B；流速：3mL/min。收集保留时间15.8min目标产物的流出液，再经氮吹干燥，加入1mL无菌生理盐水稀释备用。 18F-5MEF-F16衰变校正后比活度为127.4±25.0GBq/μmol，放化纯度如图3所示>98％。
实施例14： 18F-6FEF-F16的放化标记
18F-6FEF-F16的放射性标记合成同实施例12，不同在于采用的是1,4-二甲基吡啶 四氟硼酸盐，使用实施例10中制备的标记前体化合物3，其投药量、反应条件和HPLC分离条件一致，收集保留时间11.5min目标产物的流出液，再经氮吹干燥，加入1mL无菌生理盐水稀释备用。 18F-6FEF-F16衰变校正后比活度为93.8±12.6GBq/μmol，放化纯度如图4所示>98％。
试验例1： 19F-F16化合物的光学性质
将上述实施例1-11制备的 19F-F16化合物分别溶于PBS溶液，将溶液稀释配成10μmol/L的浓度，装于石英比色皿中，使用岛津UV-2600i分光光度计测定其吸收光谱，计算最大吸收波长；将上述测定好吸收光谱样品置于日立F-4500荧光光谱仪中，分别以其最大吸收波长为激发波长，测定其荧光发射光谱，计算最大发射波长； 19F-F16化合物荧光量子产率测定参照文献(Adv.Funct.Mater.2014,24(5),635–643)，以4-(二氰基亚甲基)-2-甲基-6-(对二甲氨基苯乙烯基)-4H-吡喃为荧光参照物，计算其在430nm下的荧光量子产率。实验结果见表1。
表1： 19F-F16化合物的光学性质表
Figure PCTCN2022136793-appb-000010
试验例2： 18F-5MEF-F16的体外稳定实验
取实施例13制备的标记产物 18F-5MEF-F16 100μCi分别加入到0.5mL的PBS溶液和胎牛血清FBS中。将两种溶液分别在37℃下分别孵育2h。在1h和2h孵育时间点，取一部分PBS样品直接进行HPLC稳定性检测。而对于FBS溶液，样品需先加入1mL乙腈使蛋白质沉淀下来，而后10000g/min离心5min，取部分上层清液，在HPLC上检测其稳定性。
实验结果如图5所示： 18F-5MEF-F16在PBS和FBS溶液中都具有较好的稳定性，没有检测到杂质产生，且放化纯度均大于98％，表明 18F-5MEF-F16在2h内是稳定的， 适于进一步体内性质探究。
试验例3： 18F-5MEF-F16在小鼠体内心肌灌注micro-PET/CT显像应用
取3只正常市售SPF级Balb/c小鼠分别经尾静脉注射100-150μCi剂量的实施例13制备的 18F-5MEF-F16探针，用2.0％的异氟烷-氧气的混合气体维持小鼠麻醉状态，注射开始同步进行PET/CT扫描(西门子Inveon microPET)，采集120min内动态显像图，勾画ROI，计算％ID/g。结果见图6和表2。
表2： 18F-5MEF-F16在正常小鼠体内摄取分布及比值
Figure PCTCN2022136793-appb-000011
如图6和表2所示： 18F-5MEF-F16在正常小鼠中的生物分布结果显示，在心肌中有较高的初始摄取值和较好的滞留。注射10min后，心肌摄取值为8.50±0.32％ID/g，注射120min后，心肌摄取值仍有7.03±0.47％ID/g，在PET/CT显像中，心脏清晰可见。探针在肾脏具有较高的摄取值，2h内呈增加趋势，表明 18F-5MEF-F16在小鼠体内主要通过肾脏排泄。 18F-5MEF-F16与非靶如肌肉比值较高，10min时心脏/肌肉为4.86，有利于提高显像对比度，获得更好的诊断效果。
试验例4： 18F-6FEF-F16在荷瘤小鼠体内micro-PET/CT显像应用
取3只市售SPF级Balb/c裸鼠，在右侧前肢腋下接种4T1细胞(小鼠乳腺癌细胞)，待肿瘤直径约为7mm，分别经尾静脉注射100-150μCi剂量的实施例14制备的 18F-6FEF-F16探针，用2.0％的异氟烷-氧气的混合气体维持小鼠麻醉状态，注射开始同步进行PET/CT扫描(西门子Inveon microPET)，采集120min内动态显像图，勾画ROI，计算％ID/g。结果见图7和表3。
表3： 18F-6FEF-F16在4T1荷瘤小鼠体内摄取分布及比值
Figure PCTCN2022136793-appb-000012
如图7和表3所示： 18F-6FEF-F16在荷瘤小鼠体内PET/CT显像表明，探针分子随时间明显浓聚于肿瘤部位，1h达到最高摄取值16.02±3.91％ID/g，明显高于肌肉、心脏、肺等脏器和组织，肿瘤/肌肉比值为7.49。PET/CT显像表明 18F-6FEF-F16能特异性靶向肿瘤部位，具有较好的靶/非靶比值，肿瘤病灶部位清晰可见，具有较好的研究及应用前景。
试验例5： 19F-F16在小鼠离体组织荧光显像应用
正常市售SPF级Balb/c小鼠分别经尾静脉注射～0.01mmol/kg剂量的实施例1-11制备的 19F-F16探针，给药2h后取小鼠离体组织或器官，经铂金埃尔默IVIS LuminaⅢ小动物活体成像仪显像观察，结果见图8。
参见图8，部分 19F-F16化合物在心脏组织分布明显，有较高靶/非靶比值，表明本发明所述化合可作为心肌灌注显像探针使用。
试验例6： 19F-EE-F16在荷瘤小鼠活体成像应用
在市售SPF级Balb/c裸鼠右侧后肢接种4T1细胞，待肿瘤直径约为7mm，经尾静脉注射～0.01mmol/kg剂量的 19F-EE-F16探针，铂金埃尔默IVIS LuminaⅢ小动物活体成像仪拍摄小鼠全身成像图，结果见图9。
参见图9，肿瘤部位能得到很好的成像效果，表明，本发明所述荧光探针在肿瘤诊断成像方面具有较好的应用前景。
以上所述仅为本发明优选实施例，对本发明而言仅具有说明性，但非限制性，相关领域普通技术人员可理解，在本发明权利要求所限定的精神和范围内可对其进行许多修改或者等效变更，但均属于本发明的保护范围。

Claims (8)

  1. 如下式I的化合物：
    Figure PCTCN2022136793-appb-100001
    其中：n为0，1，2，3或4；
    X -为阴离子；
    R 1和R 2各自独立地选自氢、卤素、氰基、硝基、C1-C6烷氧基、-NR 4R 5，其中，R 4和R 5各自独立地选自H、C1-C6烷基，或者R 4和R 5与相连的N共同构成取代或未取代的5-7元饱和杂环基，所述取代的取代基选自C1-C6烷基、C1-C6烷氧基，所述5-7元饱和杂环基含有1或2个选自N、O、S的杂原子；
    R 3为 18F或 19F。
  2. 根据权利要求1所述的式I的化合物，其中，
    X -为I -，Br -，BF 4 -或ClO 4 -；
    R 1和R 2各自独立地选自氢、卤素、氰基、硝基、C1-C6烷氧基、-NR 4R 5，其中，-NR 4R 5选自二(C1-C6烷基)氨基、吡咯基、哌啶基、未取代或C1-C6烷基取代的哌嗪基、吗啉基。
  3. 根据权利要求1所述的式I的化合物，其中，
    R 1和R 2各自独立地为：
    Figure PCTCN2022136793-appb-100002
  4. 根据权利要求1所述的式I的化合物，其中，式I化合物选自以下化合物：
    Figure PCTCN2022136793-appb-100003
    Figure PCTCN2022136793-appb-100004
  5. 根据权利要求1至4中任一项所述的式I化合物的制备方法，其制备路线如下：
    Figure PCTCN2022136793-appb-100005
    所述方法通过如下方法一或方法二进行，
    方法一
    包括如下步骤：
    (a)吲哚3-甲醛化合物1和Hal-(CH 2CH 2O) nC 2H 5OH发生亲核取代反应得到化合物2；
    (b)化合物2与对甲苯磺酰氯发生酯化反应得到化合物3；
    (c)化合物3在氨基聚醚(K2.2.2)存在下与 18F-F -发生亲核取代反应得到 18F取代的化合物 18F-4；
    (d)化合物 18F-4与1,4-二甲基吡啶盐发生Knoevenagel缩合反应得到终产物 18F-F16；
    方法二
    包括如下步骤：
    (a)吲哚3-甲醛化合物1和Hal-(CH 2CH 2O) nC 2H 5OH发生亲核取代反应得到化合物2；
    (b)化合物2与对甲苯磺酰氯发生酯化反应得到化合物3；
    (e)化合物3与四丁基氟化铵发生亲核取代反应得到 19F取代的化合物 19F-4，
    (f)化合物 19F-4与1,4-二甲基吡啶盐发生Knoevenagel缩合反应得到终产物 19F-F16；
    其中n，R 1，R 2，X -分别如权利要求1-4中所定义，Hal表示卤素。
  6. 根据权利要求1至4中任一项所述的式I化合物作为线粒体靶向的正电子发射或荧光探针的应用。
  7. 根据权利要求6所述的应用，其中所述应用为所述的式I化合物作为线粒体靶向 的正电子发射或荧光探针在心肌灌注和肿瘤显像中的应用。
  8. 根据权利要求6所述的应用，其中所述应用为所述的式I化合物作为线粒体靶向的正电子发射或荧光探针在制备心肌灌注PET显像剂或肿瘤PET显像剂或者在制备心肌灌注荧光显像剂或肿瘤荧光显像剂中的应用。
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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150336993A1 (en) * 2014-05-26 2015-11-26 The Board Of Trustees Of The Leland Stanford Junior University Mitochondria-targeted theranostic agents
CN109293633A (zh) * 2018-10-26 2019-02-01 山东大学 一种非反应型线粒体追踪荧光探针ivpi-12及其应用
CN109867591A (zh) * 2019-03-12 2019-06-11 浙江大学 18f标记的aie荧光/pet双模态探针及其制备方法和应用
CN110577478A (zh) * 2018-10-25 2019-12-17 南方医科大学南方医院 一种正电子探针及其制备方法与应用
WO2020150103A1 (en) * 2019-01-15 2020-07-23 The Board Of Trustees Of The Leland Stanford Junior University Delocalized lipophilic cation compounds and methods of use thereof
CN112028815A (zh) * 2019-06-03 2020-12-04 中国药科大学 吲哚类衍生物及其医药用途
CN112375027A (zh) * 2020-12-07 2021-02-19 中国药科大学 吲哚磺酰胺类衍生物及其医药用途

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150336993A1 (en) * 2014-05-26 2015-11-26 The Board Of Trustees Of The Leland Stanford Junior University Mitochondria-targeted theranostic agents
CN110577478A (zh) * 2018-10-25 2019-12-17 南方医科大学南方医院 一种正电子探针及其制备方法与应用
CN109293633A (zh) * 2018-10-26 2019-02-01 山东大学 一种非反应型线粒体追踪荧光探针ivpi-12及其应用
WO2020150103A1 (en) * 2019-01-15 2020-07-23 The Board Of Trustees Of The Leland Stanford Junior University Delocalized lipophilic cation compounds and methods of use thereof
CN109867591A (zh) * 2019-03-12 2019-06-11 浙江大学 18f标记的aie荧光/pet双模态探针及其制备方法和应用
CN112028815A (zh) * 2019-06-03 2020-12-04 中国药科大学 吲哚类衍生物及其医药用途
CN112375027A (zh) * 2020-12-07 2021-02-19 中国药科大学 吲哚磺酰胺类衍生物及其医药用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ZHENG LINGLING, WANG ZHIMING, ZHANG XIAOQING, ZHOU YUJING, JI AIYAN, LOU HONGYUE, LIU XINGDANG, CHEN HAO, CHENG ZHEN: "Development of Mitochondria-Targeted Small-Molecule Dyes for Myocardial PET and Fluorescence Bimodal Imaging", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 65, no. 1, 13 January 2022 (2022-01-13), US , pages 497 - 506, XP093072195, ISSN: 0022-2623, DOI: 10.1021/acs.jmedchem.1c01660 *

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