Tau蛋白显像剂~(18)F-THK5317的合成与初步临床应用

~(18)F-THK5317是以tau为靶点的新型分子探针,本研究利用国产氟多功能模块自动化合成~(18)F-THK5317,在动物实验基础上进行了初步的临床研究。以(S)-2-(4-甲氨基苯基)-6-[[2-(四氢吡喃基-)-3-对甲苯磺酰氧基]丙氧基]喹啉为前体,经亲核反应、酸水解、碱中和,分别采用混合液直接HPLC纯化与混合液经C18小柱预纯化后再HPLC分离纯化两种方法得到~(18)F-THK5317;研究了药物在正常KM小鼠体内生物学分布;对比了~(18)F-THK5317在正常人(HC)和阿尔茨海默病(AD)患者脑中PET/MR显像结果。先以C18小柱预纯化粗产品再用HPLC分离,能显著改善HPLC分离效果和提高产品放化纯度。~(18)F-THK5317未校正合成产率为(18.7±5.3)%(n=7),放化纯度大于95%。小鼠生物分布表明,探针易穿透血脑屏障,并且能迅速从正常脑组织清除,Brain_(1 min)/Brain_(60 min)放射性摄取比为34;PET/MR结果显示,AD患者双侧颞叶、皮层的放射性滞留均高于健康对照。以上结果表明,国产氟多功能模块能够稳定高效地合成符合药物质控标准的~(18)F-THK5317,动物实验及初步临床研究表明~(18)F-THK5317具有在体显像tau蛋白的潜力。     

在线合成心肌灌注显像剂:碳-11三苯甲基磷

99mTc-MIBI是常用的心肌灌注显像剂,肝摄取高,临床应用有一定的影响。11C-三苯甲基磷(11C-TPMP)为脂溶性正一价阳离子,用于心肌显像能得到更好的分辨率。以11CH3-Triflate为甲基化试剂,采用LOOP法在线合成了11C-TPMP,研究了11C-TPMP在正常小鼠的分布,正常大鼠行Micro PET显像,并与4位18F-苯基三苯基磷(18F-FTPP)比较。结果显示,LOOP法合成11C-TPMP效率大于90%(校正),放化纯度大于98%。正常小鼠心肌有高的摄取,注射后5 min和60 min,心肌摄取分别为(71.9±19.9)%ID.g–1和(12.87±0.81)%ID.g–1,肝摄取低,注射后5 min和60 min心肝摄取比分别为11.57和3.2。正常大鼠micro PET显像显示,从20 min到60 min,心肌显像清晰,肝未显像。表明LOOP法可高效合成11C-TPMP,11C-TPMP是一个很有希望的心肌灌注显像剂。     

Tau蛋白显像剂18F-THK5317的合成与初步临床应用

¹⁸F-THK5317是以tau为靶点的新型分子探针,本研究利用国产氟多功能模块自动化合成¹⁸F-THK5317,在动物实验基础上进行了初步的临床研究。以(S)-2-(4-甲氨基苯基)-6-［［2-(四氢吡喃基-)-3-对甲苯磺酰氧基］丙氧基］喹啉为前体,经亲核反应、酸水解、碱中和,分别采用混合液直接HPLC纯化与混合液经C18小柱预纯化后再HPLC分离纯化两种方法得到¹⁸F-THK5317;研究了药物在正常KM小鼠体内生物学分布;对比了¹⁸F-THK5317在正常人（HC）和阿尔茨海默病（AD）患者脑中PET/MR显像结果。先以C18小柱预纯化粗产品再用HPLC分离,能显著改善HPLC分离效果和提高产品放化纯度。¹⁸F-THK5317未校正合成产率为（18.7±5.3）%（n=7）,放化纯度大于95%。小鼠生物分布表明,探针易穿透血脑屏障,并且能迅速从正常脑组织清除,Brain_{1 min}/Brain_{60 min}放射性摄取比为34;PET/MR结果显示,AD患者双侧颞叶、皮层的放射性滞留均高于健康对照。以上结果表明,国产氟多功能模块能够稳定高效地合成符合药物质控标准的¹⁸F-THK5317,动物实验及初步临床研究表明¹⁸F-THK5317具有在体显像tau蛋白的潜力。     

O-（2-[^18F]氟代乙基）-5-羟基-L-色氨酸的合成、生物分布及MicroPET显像

通过对现有CTI公司计算机控制的化学合成模块（CPCU）进行改造,合成了L-5-羟基色氨酸类似物（5-^18FEHTP）,并用高效液相（HPLC）检测其放化纯度,所得产品用于昆明小鼠的S180肉瘤模型显像。结果显示,采用改进方法合成5-^18FEHTP的总时间是45min,放化收率为12％～16％（n=15）,产品的放化纯度〉98％。正常昆明小鼠体内生物分布显示,其脑组织摄取较低,最大摄取率为（2．354±0．405）％ID／g。血液清除较快,30min时摄取率已降为（2．974±0．278）％ID／g。肾脏给药60min摄取率值最大为（11．706±0．374）％ID／g,5-^18FEHTP经过肾脏排出体外。MicroPET小鼠显像结果表明,5-^18FEHTP在S180肉瘤中浓集程度明显高于周围其它组织。以上结果提示,利用CPCU半自动合成5-^18FEHTP,方法简便、稳定,产品纯度较高。小鼠生物分布和显像结果表明,5-^18FEHTP可能成为一种新的PET显像剂。     

国产氟多功能模块自动化合成~(18)F-6-氟-L-DOPA及其临床初步应用

~(18)F-6-氟-L-DOPA(~(18)F-DOPA)在神经内分泌肿瘤诊断中具有重要价值,本研究利用国产氟多功能模块合成了~(18)F-DOPA并进行临床初步研究。以6-硼酯-二甲氧基-L-DOPA为前体,铜盐Cu(OTf)2(py)4为催化剂直接亲核反应,再经氢碘酸水解得到~(18)F-DOPA,并进行质量控制,测量其体外稳定性,研究~(18)FDOPA在正常人和一例神经内分泌肿瘤患者的显像。~(18)F-DOPA合成耗时60min,不校正合成效率为(10.0±2.3)%(n=6),放化纯度大于99%,产量大于7.4GBq;加入微量抗坏血酸或乙醇可提高其体外稳定性。人体PET结果显示:~(18)F-DOPA在脑内浓聚于纹状体,主要经肾脏代谢,见胰岛细胞瘤显像。结果表明,国产氟多功能模块自动化合成~(18)F-DOPA简单快捷、重复性好,产品质量符合临床要求。     

~(18)F-DCFPyL的制备和初步临床应用

利用国产氟多功能模块自动化合成前列腺特异性膜抗原(PSMA)显像剂~(18)F-DCFPyL(2-(3-{1-羧基-5-[(6-[~(18)F]氟-吡啶-3-羰基)-胺基]-戊基}-脲基)-戊二酸)并进行生物学评价。通过改变不同的水解条件,研究氟化中间体的水解效率,对~(18)F-DCFPyL进行质量控制和生物安全性评价,并通过小鼠生物学分布和初步临床试验探讨~(18)F-DCFPyL的生物学性质,一例生化复发的前列腺癌术后患者进行了~(18)FDCFPyL PET/CT显像。结果表明,H3PO4、HCl和HI三种酸的水解效率相近,分别为17.1%、16.9%和18.4%,得到产品放化纯度均大于98%,比活度为54~90GBq/μmol,~(18)F-DCFPyL在小鼠体内经泌尿系统排泄,其余组织无明显摄取,前列腺癌术后生化复发的患者~(18)F-DCFPyL PET显像能够探查同机CT无法检出的微小转移灶。国产氟多功能模块自动化合成~(18)F-DCFPyL的稳定性和重复性高,产品质量符合临床研究要求,~(18)F-DCFPyL具有良好的生物学分布特性,可用于早期诊断前列腺癌及探测前列腺癌生化复发患者的病灶。     

氟［18F］比他班自动化合成及初步临床应用

氟［¹⁸F］比他班（¹⁸F-florbetaben）是美国FDA于2014年批准上市的β-淀粉样蛋白显像剂,主要用于诊断阿尔茨海默病（AD）或其他认知障碍疾病。本研究使用改良后的国产氟多功能模块,建立¹⁸F-florbetaben自动化生产工艺,并针对其临床应用效果进行初步验证。结果显示,¹⁸F-florbetaben自动化合成耗时38 min,不校正合成效率为（45.0±2.3）%（n=6）,放化纯度大于95%,其临床PET显像效果理想。结果表明,国产氟多功能模块可实现¹⁸F-florbetaben的自动化生产,且工艺可靠,合成时间短。本文研究成果有助于推动该显像剂的国内临床使用。     

国产氟多功能模块合成雌激素受体显像剂16α-[18F]氟-17β-雌二醇

采用国产氟多功能模块,以3-甲氧基甲基-16,17-O-磺酰基-表雌三醇-O-环状砜（3-O-(Methoxymethyl) -16,17-O-sulfuryl-16-epiestriol,MMSE）为前体,在国产氟多功能合成模块的密封体系下,经¹⁸F标记合成雌激素受体显像剂16α-[¹⁸F]氟-17β-雌二醇（¹⁸F-FES）。结果显示：合成的¹⁸F-FES,不校正合成效率为8.2%,校正合成效率为12.8%;合成时间约为70 min,标记物¹⁸F-FES放化纯度大于98%,体外稳定性良好。以上结果表明,国产氟多功能模块可制备¹⁸F-FES溶液,制备的¹⁸F-FES溶液符合放射性药物的质量要求。     

柱分离法自动化合成~(18)F-FLT

使用PET-MF-2V-IT-I型氟-18多功能合成模块,以3-N-t-叔丁氧羰基-1-[5’-O-(4,4’-二甲氧基三苯甲基)-2’-脱氧-3’-O-(4-硝基苯磺酰基-β-1)-苏戊呋喃糖]胸腺嘧啶为前体,经氟化、水解后小柱分离制得18F-FLT注射液。对荷肝细胞癌小鼠进行18F-FLTPET/CT显像,结果显示,柱分离法合成18F-FLT耗时～35min,放化产率为12%-15%,放化纯度95%。表明18F-FLT静脉注射1h后肿瘤对18F-FLT的摄取明显高于周围正常组织。该法合成简单、反应时间短、产率高,可满足临床应用。     

肿瘤乏氧显像剂18F-FAZA的自动化合成

为了制备1-α-D-[5,-脱氧-5’-氟阿拉伯呋喃糖基]-2-硝基咪唑(18FAZA),采用两锅法与氟多功能模块,以4～6 mg前体在115℃氟化反应10 min,用C-18柱捕获中间产物,用NaOH水解,HCl中和,进行HPLC分离,得到18FAZA注射液体.总合成时间45 min,放化产率和放射化学纯度分别大于1...     

Preparation andBiodistribution of Myocardial Perfusion Imaging Agent 18F-TPT

⁹⁹Tc^m-sestamibi is typically used as amyocardial perfusion imaging agent for SPECT, however, the high uptake of liverand lung compromise the diagnostic accuracy. PET has higher spatial resolutionand quantitative measurement of myocardial tracer uptake. The lipophiliccationic compound, (4-[¹⁸F]fluorophenyl)triphenylphosphoniumion (¹⁸F-TPT)was synthesized as a potential positron emission tomography (PET) myocardialperfusion agent, biodistribution studies in the NH rats and Micro PET/CTimaging studies in the SD rats were performed. Total synthesis time was about 1h and the uncorrected synthesis yield was 2.5%, radiochemical purity was higherthan 99.5%, the product had good stability at room temperature. Biodistributiondata in rats showed high levels of accumulation in the heart with stableretention and rapid blood clearance, Heart-to-liver ratios at 30, 60, 90,and120 min were 33.1, 14.8, 25.7 and 17.3, respectively; Micro PET/CT imagingin the SD rat showed intense cardiac uptake and non-target tissues as liver,lung uptake were washed out quickly. The result show that ¹⁸F-TPT may have potential as amyocardial perfusion imaging agent for PET.     

(N-[~(18)F]氟甲基)-胆碱的自动化合成及其生物分布研究

采用住友CFN-multi-P100多功能模块快速、自动化合成(N-[~(18)F]氟甲基)-胆碱(~(18)F-FCH),并评价其在正常小鼠体内生物分布,以及胰腺癌裸鼠模型的PET/CT显像情况。前体CH2Br2与~(18)F-气相反应生成18FCH2Br,18FCH2Br经4个Si柱纯化后与三氟甲基磺酰银(Ag-Triflate)反应生成活性更高的氟代三氟甲基磺酰基甲烷(~(18)FCH2OTf),新中间体与预先加在C-18柱子上的N,N-二甲基乙醇胺(DMAE)反应再经SEP-PAK CM柱纯化得到18F-FCH。将~(18)F-FCH静脉给予正常小鼠,分别在给药后5、10、30、60、90、120min处死,测定主要脏器的质量及放射性计数。将~(18)F-FCH静脉给予胰腺癌裸鼠,注射10min后观察荷瘤裸鼠的PET/CT显像情况。结果显示,~(18)F-FCH合成时间32min,未校正的合成效率为(25±5)%(n=23),放化纯度大于97%。小鼠体内生物分布实验显示,18F-FCH在血液中清除快,绝大多数脏器在5min时放射性分布达最高值,后逐渐降低或处于相对稳定状态。放射性主要分布在肾脏、肝脏,而脑、肺、肌肉对~(18)F-FCH的摄取均较少。荷瘤(胰腺癌)裸鼠的PET/CT显像表明,~(18)FFCH在裸鼠肾脏、肝脏和脾脏聚集,胰腺癌细胞对~(18)F-FCH未见明显摄取。结果提示,住友CFN多功能模块可自动化、快速合成18F-FCH。18F-FCH在正常小鼠体内分布与文献报道的11 C-胆碱相似,具有一定的应用前景,但其对胰腺癌的诊断仍需进一步研究。     

正电子心肌灌注显像剂的临床应用及研究进展

随着正电子发射型断层显像仪（PET）及PET／CT在国内外的逐步推广应用,正电子心肌灌注显像剂的研究也备受关注,PET应用的心肌灌注显像剂有^82Rb ^13NH3和H2^16O,但半衰期均较短（t1/2〈10min）,或需要^82Sr／^82Rb发生器或加速器等原因限制了其应用。近来长半衰期核素^18F标记的新型灌注类显像剂成为研究热点,报道较多的一类显像剂是以心肌细胞线粒体复合物-I（MC—I）为靶点,该类显像在动物体内外试验中均表现出良好的显像性能：心肌摄取快,血流灌注相关性好,滞留时间长。其中又以^18F-BMS747158—02最为突出,除具备上述特性外,还体现出心室放射性摄取均匀,体内生物分布理想的特点,有望成为较理想的正电子心肌灌注显像剂。     

Aβ显像剂18F-AV45的自动化合成及临床验证

使用进口氟多功能合成模块TRACERlab FX2 N合成器自动化合成β-淀粉样蛋白（β-amyloid protein, Aβ）正电子显像剂¹⁸F-AV45,并进行临床验证。在TRACERlab FX2 N合成器上,以AV105为前体,与18F-发生亲核反应后,依次经酸水解及碱中和,经过高效液相色谱法（high performance liquid chromatography, HPLC）分离并纯化后获得¹⁸F-AV45,进行质量控制。并用制备的¹⁸F-AV45对1例阿尔兹海默病（Alzheimer disease, AD）患者及1例健康对照者行¹⁸F-AV45 PET/CT扫描。结果表明,¹⁸F-AV45合成时间为80 min,不校正合成效率为（17.02±1.52）%（n=6）,产品放化纯度大于95%。临床应用显示¹⁸F-AV45在AD患者大脑皮层摄取弥漫增高,提示大脑皮层β淀粉样蛋白沉积;在健康对照者大脑皮层未见明显摄取,即大脑皮层未见β淀粉样蛋白沉积。TRACERlab FX2 N合成器自动化合成¹⁸F-AV45简便快捷,重复性好,制备出的¹⁸F-AV45产品质量符合临床要求,该合成方法可为¹⁸F-AV45模块合成提供参考。     

基于国产氟多功能合成模块的~(18)F-FMISO自动合成工艺验证

为验证特异性肿瘤PET乏氧显像剂1-H-1-(3-~(18)F-2-羟基丙基)-2-硝基咪唑(~(18)F-FMISO)注射液的临床前即时标记工艺的可行性、可靠性和稳定性,采用国产氟多功能自动化合成装置,以1-(2′-硝基-1′-咪唑基)-2-O-四氢吡喃基-3-O-甲苯磺酰基丙二醇(NITTP)为前体,经氟化、水解反应制备18 F-FMISO注射液,按照优化的制备工艺进行~(18)F-FMISO三批连续生产,并对其关键工艺参数和产品质量标准进行验证。结果表明:总合成时间小于40min,产品放化产率大于45%(未衰减校正,n=5),比活度大于3.7×1010 Bq/mmol,放置3个半衰期后放化纯度仍大于95%,体外稳定性良好。该自动合成工艺稳定可行,三批产品各项指标均符合质量标准规定,满足临床PET显像要求。     

5-羟色胺转运蛋白显像剂~(11)C-DASB的自动化合成及Micro PET/CT显像

目的:自动化合成5-羟色胺转运蛋白显像剂11 C-DASB并进行大鼠Micro PET/CT显像;方法:通过改变甲基化试剂、溶解前体溶剂及反应条件,得到优化的标记条件作为碳-11多功能合成模块的输入参数,进行自动化合成11 C-DASB,大鼠静脉注射11 C-DASB 45 min后进行显像;结果:采用11 C-CH3-Triflate作为甲基化试剂,通入新配制的含1mg去甲基DASB前体的500μL DMSO溶液内,80℃下加热2min,标准率为63.7%,大鼠显像表明,11 C-DASB特异性的浓聚于SERT富集区域;结论:经优化,11 C-DASB自动化合成可得到较高产率,大鼠显像表明,其特异性浓聚于SERT富集区域,有望作为5-羟色胺转运蛋白显像剂。     

Distribution of Dopamine D2 Receptor PET Imaging Agent 18F-Fallypride in the Brain of Normal and Aged Rats

With the progress of population aging, the incidence of age related disease has greatly increased. The dopamine D2 receptor is closely related to the age-related diseases, such as PD and AD. The PET imaging of the dopamine D2 receptor can provide noninvasive, dynamic, early and quantitative information on the function of the brain. So we intend to prepare dopamine D2 receptor PET imaging agent¹⁸F-Fallypride and to study the disturbution of the agent in the brain of normal and aged rats, further to explore the relationship between dopamine D2 receptor and senility.¹⁸F-Fallypride was prepared by nucleophilic reaction. And the PET image was performed in aged and normal rats 15 minutes after injection of the agent .Striatums were delineated as the region of interesting (ROI), the standard uptake value (SUV) of which was calculated. PMOD was used for image fusion ,partition and quantitative analysis of standard uptake values of each brain area. After imaging, cardiac ventricle was perfused. The brain was obtained and frozen. The contour structure was observed by HE staining.The results showed that the labeling yield was over 95% and the radiochemical purity was higher than 98%. The stability was still over 95% 2 hours after incubation with PBS. The striatum uptake of¹⁸F-Fallypride radioactivity in aged and normal rats 15 minutes after injection were (0.58±0.11)%ID/g, (0.39±0.14)%ID/g. The uptake of cortex cingulate, cortex insular, hypothalamus, olfactory, midbrain in normal rats ((0.120±0.012)%ID/g, (0.182±0.002)%ID/g, (0.111±0.002)%ID/g, (0.127±0.007)%ID/g, (0.083±0.012)%ID/g respectively) were inferior to aged rats ((0.154±0.013)%ID/g, （0.344±0.014）%ID/g, （0.244±0.019）%ID/g, （0.263±0.020）%ID/g, （0.216±0.012）%ID/g), which was displayed by PMOD. HE staining showed severe brain injury in elderly SD rats. Some neurons in the aged SD rats appeared acidophil change or nuclear fragmentation, accompanied by spongy deformation, lamellar or focal neuronal necrosis, and no obvious morphological changes. The relevance between dopamine D2 receptor and senility was demonstrated by PET imaging, which provided a basis for further research on the methodology of disease and pharmacodynamics research.     

Full Automated Synthesis of18F-FDOPA and Preliminary PET/CT Imaging

¹⁸F-fluoro-L-dihydroxyphenylalanine (¹⁸F-FDOPA) as a dopamine neurotransmitter imaging agent has been widely used for diagnosis and therapy evaluation of Parkinson's disease, brain tumors and neuroendocrine diseases with positron emission tomography (PET) imaging in clinical setting and research. To meet the increasing clinical demand in oncology and neurology, a routine protocol for the automated synthesis of¹⁸F-FDOPA with a disposable cassette system on an imported multifunctional synthesizer was studied and discussed.¹⁸F-FDOPA was automatically synthesized via a multiple-step reaction, including fluorination, reduction, iodization alkylation and hydrolysis, following purification by using a semi-preparative high-performance liquid chromatography (HPLC) system which was built in the multifunctional synthesizer. After HPLC purification, the purified¹⁸F-FDOPA solution was collected and passed through a sterilizing filter into a collection bottle. The final¹⁸F-FDOPA injection was obtained for quality control (QC) determination. The QC indexes of the final products were detected： the injection was colorless and transparent, pH value was at 4 to 5.5, radiochemical purity >98%, radionuclide purity >99%, specific activity >1.9 GBq/μmol, K_2.2.2 content <50 mg/L, methanol content <0.01%, alcohol content <0.01%, dichloromethane content <0.01 mg/L, dimethylformamide content <15 mg/L, bacterial endotoxin test <0.100 EU/mL, sterility test 0 cfu/mL,and abnormal toxicity test was negative. PET/CT imaging of rats was performed by intravenous injection of¹⁸F-FDOPA half an hour after the intraperitoneal injection of carbidopa, PET/CT scan was performed after 100 min post-injection. The imaging of¹⁸F-FDOPA showed symmetry high uptake in the bilateral striatum of normal rats. The decay-corrected radiochemical yield of¹⁸F-FDOPA from the¹⁸F-fluoride was (63.1±3.8)% (n=10) at the end of synthesis (EOS), the radiochemical purity was no less than 98%, and the total radiosynthesis time was within 80 min. The quality control results demonstrated that the quality indexes of the final injection solution met the relevant requirements of radiopharmaceutlcals, which were well-suited for clinical application. An efficient and high reproducible automatic method for the radiosynthesis of¹⁸F-FDOPA with high radiochemical yields and good radiochemical purity is obtained and performed via a multi-step reaction on the multifunctional synthesizer.¹⁸F-FDOPA can be used for animal and human PET imaging.     

Preparation and Clinical Application of 18F-DCFPyL

¹⁸F-DCFPyL, a PSMA-based PET imaging agent for prostate cancer, was auto synthesized and evaluated. Following the direct nucleophilic heteroaromatic substitution with ［¹⁸F］fluoride at the ortho-position of precursor, the deprotection of the ester moieties of the intermediate with different acids was attempted to obtain a good hydrolysis yield. The biodistribution in normal NIH mice and PET/CT imaging for a patient with biochemical recurrence of prostate cancer were also performed. The results showed that no remarkable discrepancy of the hydrolysis efficiency was found among three kinds of acids, H₃PO₄, HCl and HI, which were 17.1%, 16.9% and 18.4%, respectively with a specific activity of 54 to 90 GBq/μmol. The highest levels of radioactivity in the NIH mice were observed in the kidneys. Meanwhile, the uptake of the tracer in the blood was declined rapidly and a low accumulation of the radio-tracer was observed in most of the other organs. ¹⁸F-DCFPyL PET imaging for a postoperative patient with biochemical recurrence of prostate cancer can detect small metastatic foci that can not be detected by the CT. ¹⁸F-DCFPyL was synthesized reliably and repeatedly by domestic synthesis module and it passed the quality control. It has satisfactory properties in vivo and is probably suitable for early diagnosis of prostate cancer and detection of lesions in patients with biochemical recurrence of prostate cancer.