多巴胺转运蛋白显像剂^18F—FP—β—CIT的制备

可卡因经水解、脱水、甲酯化、苯基溴镁格氏反应、脱N－甲基、碘化、N烷基化、甲磺酰化等反应制备氟标记前体,用K222催化进行氟标记,得到^18F-FP-β-CIT[N－（3－氟丙基）－2β－甲酯基－3β－（4‘－碘苯）去甲基托烷],标记率25％－30％,放射化学产率10％－12％,合成及纯化时间100－110min,纯化后放化纯度＜95％,稳定性较好。     

多巴胺转运蛋白显像剂β-CIT的合成及其碘标记

报道了β－ＣＩＴ（２β－甲酯基－３β－（４’－磺苯基）托烷）及标记前体（２β－甲酯基－３β－（４’－（三丁基锡）苯基）托烷）的合成路线，化合物ＩＲ、ＮＭＲ、ＭＳ、元素分析等数据与结构相符，由标记前体制备＾１２５／１３１Ｉ－β－ＣＩＴ，标记率和放化纯度均〉９０％。     

多巴胺转运蛋白显像剂FP-β-CIT的合成及碘标记

以可卡因为原料经水解、脱水、甲酯化、苯基溴镁格氏反应、脱N-甲基、碘化、N烷基化，烷基锡化等反应制备FP-β-CIT标记前体。用过氧乙酸法进行碘标记，得到^131I-FP-β-CIT。标记率及放射化学纯度高（大于95%），稳定性好。     

β-CIT的131I标记及其初步临床应用

用过氧乙酸法进行了β-CIT的131I标记,并用标记物对4例正常对照、8例帕金森氏病(PD)患者、3例帕金森氏综合征(PS)患者进行显像,计算纹状体与小脑的放射性摄取比(特异性摄取)及纹状体131I-β-CIT摄取的非对称指数AI.结果显示131I-β-CIT放化纯度达(97.6±0.3)%,室温下放置4 h以及分别与水、人新鲜血清孵育4 h后,其放化纯度仍均＞95%;纹状体能特异摄取131I-β-CIT,与正常对照组及PS组相比,PD患者双侧纹状体摄取的放射性明显降低(P＜0.01),且症状对侧降低更明显;PS患者与正常对照组相比,其纹状体摄取无明显差异(P＞0.05).直线回归分析显示,PD患者症状的严重程度与纹状体特异摄取131I-β-CIT下降密切相关.提示131I-β-CIT多巴胺转运蛋白显像可作为PD诊断、鉴别诊断和病情严重程度的客观指标.     

^131I标记肿瘤血管靶向多肽的实验研究

设计合成含精氨酸-精氨酸-亮氨酸（RRL）序列的多肽，并用氯胺-T法对其进行^131I标记，标记率约60％，放化纯度〉99％。体外放置24h放化纯度仍≥90％。肿瘤对^131I标记多肽的摄取在注射后明显高于其他器官，注药后24h，肿瘤对^131I-多肽的摄取约为0．65％ID／g，肿瘤与肌肉的放射性摄取比（T／NT）达9．08。以上结果表明：通过氯胺-T法对该多肽进行^131I标记是可行的，标记物可在体外稳定存放24h；^131I-多肽可以靶向肿瘤血管，有进一步研究的价值。     

多巴胺转运蛋白显像剂131I-FP-β-CIT的临床前药理研究

报道了用自制的^131I—FP—β—CIT(N-（3’-氟丙基)-2β-羰甲氧基-3β-(4’-碘苯基)托烷)测定分配比，进行大鼠体内及脑内分布、兔血药清除动力学、大鼠脑放射自显影、猴显像和异常毒性等实验。结果显示：^131I—FP-β—CIT脂溶性大，在pH为7．0和7.4时的分配比分别为15和28；药物能迅速进脑，并有较好的滞留(2min时脑摄取为0．78％ID，2h时为O．59％ID)；脑内药物在纹状体中浓聚，1h时纹状体与小脑、额叶、海马的比值分别为5．23、2．15和3．10；纹状体的摄取能被B—CFT阻滞，表明药物与多巴胺转运蛋白(DAIT)结合的亲和性和特异性较好；药物在兔血中清除迅速，血药浓度降至一半所需时间约为2min；放射自显影显示出药物在纹状体区域的放射性浓聚，左右纹状体基本对称，纹状体与顶叶的比值为2．7；正常猴显像表明在注药后2h时纹状体与颞叶、小脑的比值分别为5．4和3．0；异常毒性实验结果显示小鼠所耐受的剂量为人的750倍，表明药物非常安全。以上结果表明^131I—FP—β—CIT与DAT有很好的亲和性与特异性，其体内性质适合显像，具有良好的临床应用前景。     

多巴胺D2受体显像剂Epidepride的合成及其^131I标记

以3－甲氧基水杨酸为原料合成了Epidepride[S-(-)-N-[1-乙基－2－吡咯烷基]-5－碘－2,3－二甲氧基基甲酰胺]及其标记前体（S－（－）－5－（三正丁基锡）－N（1－乙基－2－吡咯烷基）甲基]2,3-二甲氧基苯甲酸酰胺）,并采用双氧水法对标记前体进行^131I标记,获得了^131I-epidepride,标记率和放化纯度均大于95％。132I-epidepride溶液体外稳定性好,4℃放置15d放化纯度仍大于90％。^131I-epidepride与D2受体亲和力高,大鼠脑内纹状体与小脑的摄取比在320min时高达237;在脑内纹状体的吸收可被Spiperone安全阻断。因此,^131I－epideride有望成为多巴胺D2受体SPECT显像剂。     

5-羟色胺与多巴胺系统相互作用的研究

２β－甲酯基－３β－（４－碘苯基）托烷（β－ＣＩＴ）与多巴胺转运蛋白（ＤＡＴ）、５－羟色胺转运蛋白（５－ＨＴＴ）亲和力几乎相等，因此可用５－ＨＴ重摄取抑制氯米帕明来研究５－ＨＴ系统与ＤＡ系统间的相互作用。用过氧乙酸氧化碘标记法制备＾１２５Ｉ－ＣＩＴ。用不同剂量盐酸氯米帕明（０．２５、０．５、１、１．５、２、２．５ｍｇ）研究对＾１２５Ｉ－β－ＣＩＴ脑内结合的影响，及同一剂量盐酸氯米帕明（１ｍｇ／０．     

新型乏氧肿瘤显像剂^99Tc^m-MNLS、^99Tcm-MLS的合成及其在小鼠体内的生物分布

合成了含有硝基咪唑基团的磷酸酯衍生物2-（2-methyl-5-nitro-1H—imidazol-1-yl）Ethyl Dihydrogen Phosphate（MNLS）和它的非硝基类似物2-（2-methyl-1H-imidazol-l-yl）Ethyl Dihydrogen Phosphate（MLS），采用^99Tc^m直接法对其进行标记，并研究了^99Tc^m-MNLS和^99Tc^m-MLS的理化性质及其在荷EMT-6小鼠体内的生物分布。采用最佳标记条件后，^99Tc^m-MNLS和^99Tc^m-MLS的标记率均〉90％。荷EMT-6小鼠体内生物分布表明：^99Tc^m-MNLs的肿瘤放射性摄取率、肿瘤与肌肉和肿瘤与肝的放射性摄取比（T／NT）（120min时分别为2．99±0．25％ID／g、5．90和1．03）显著高于已知的乏氧显像剂^99Tc^m-HL91（120min时分别为0．93±0．13％ID／g、3．59和0．17）和不含硝基基团的类似物^99Tc^m-MLS（120min时分别为1．61±0．13％ID／g、5．40和0．13）。与^99Tc^m-MLS相比^99Tc^m-MNLS配体中的硝基对于肿瘤的摄取影响很大，这表明^99Tc^m-MNLS的肿瘤摄取与其乏氧机制有关。上述研究结果表明，^99Tc^m-MNLS具有肝摄取低，肿瘤摄取较高的优点，作为潜在的新型乏氧肿瘤显像剂，值得进一步研究。     

^123I—β—CIT：一种新型多巴胺转运体显像剂

^123I-β-CIT是一种新型多巴胺转运体SPECT显像剂,具有亲和力高,脑滞留时间长,以及纹状体与小脑摄取比较高等特点。它对帕金森氏病的早期诊断、临床分期、疗效观察有较高的临床应用价值。本文介绍了^123I-β-CIT在标记、药物动力学、毒性及临床应用等方面的研究结果,并提出了今后在DAT显像剂方面的发展方向。     

The preclinical pharmacological study of dopamine transporter imaging agent ^18F-FP-β-CIT

The paper is to study pharmacologic characteristics of 18F-FP-β-CIT (18F-N-(3-fluoropropyl)-2β-carbomethoxy-3β- (4-iodophenyl)nortropane) as an imaging agent for dopamine transporter. The radiochemical purity of 18F-FP-β-CIT in aqueous solution was over 95% after standing at room temperature for 4h. Biodistribution displayed rapid uptake in rat brain (1.375 %ID/organ at 5min and 0.100 %ID/organ at 180 min) and the striatal uptake was 1.444, 0.731, 0.397, 0.230 and 0.146 %ID/g at 5, 30, 60, 120 and 180 min, respectively. The values of striatum/cerebellum, striatum /frontal cortex and striatum / hippocampus in rat's brain at 30 min were 3.38, 2.17 and 2.40 respectively. The uptake in striatum can be blocked by β-CFT, suggesting that 18F-FP-β-CIT binds to DAT peculiarly. The compound was rapidly cleared from monkey's blood. The striatal uptake was bilaterally decreased in the left-sided lesioned PD rats, compared with normal control. Brain PET imaging studies in normal monkey showed that 18F-FP-β-CIT was concentrated in striatum. The test of undue toxicity showed that the dose received by mice was 1250 times as by human, which indicates that 18F-FP-β-CIT is very safe. So 18F-FP-β-CIT is a promising PET imaging agent for DAT with safety and validity.     

Pharmacological studies of dopamine transporter imaging agent 125／131I－β－CIT

To prepare ^125/131I-β-CIT(2β-carbomethoxy-3β-(4-iodophenyl)tropane) as an imaging agent for dopamine transporter (DAT),the labeling method from tributylstannyl precursor with peracetic acid has been reported in this article.The radiochemical purity(RCP) of the labeled compound was over 95% determined by HPLC and TLC.The stability,partition coefficients were also determined.The pharmacological studies of the imaging agent were performed in rats,mice,rabbits and normal monkey.The ligand showed preferable uptake in rats,mice,rabbits and normal monkey.The ligand showed preferable uptake in brain (1.9%ID/organ in rats and 4.5%ID/organ in mice at 5min).The ratios of striatum/cerebellum,hippocampus/cerebellum and cortex/cerebellum were 29.8,3.97and 4.75 at 6h in rats,and 8.52,2.99 and 3.06 at 6h in mice,respectively.In monkey brain imaging the ratios of striatum/frontal cortex(ST/FC)and striatum/occipital cortex(ST/OC) were 5.14 and 5.97 at 4h.respectively,All of above showed the high affinity of the ligand to DAT,The compound was primarily metabolized in liver because the hepatic uptake was much higher than other organs(75.4%ID/organ at 18h).The half-life of blood elimination was 5min.The dose received by mice was 2500 times as high as that received by human in the test of undue toxicity,which evaluated the safety of the agent.All the results suggest that β-CIT can be used as a potential DAT imaging agent.     

Synthesis of [~(18)F]-N-3-fluoropropyl2β-carbomethoxy-3β-(4-iodophenyl)nortropane ([~(18)F]-FP-β- CIT)

1 INTRODUCTIONSingle photon emission computed tomography (SPECT) and positron emission tomography (PET) provide sensitive and powerful means for detecting specific moleculartargets in brain. In neuron abnormality, molecular targets for SPECT or PET brainimaging agents may be used to reveal the status of associated neurons. The dopaAnne(DA) transporter (DAT) is a protein complex locallied ajmost exclusively presynaptically at the dopalninergic nerve terminal. Increasing evidence s…     

Radioiodine-Labeling of Chlorpyrifos and Its Biodistribution in Mice

To investigate the preparation of radioiodinated Chlorpyrifos and its biodistribution in mice, Chlorpyrifos was labeled with¹³¹I using the Iodogen method. Biodistribution studies were carried out in KM mice. At different times after radiopharmaceutical i.v. administration (185 kBq¹³¹I-Chlorpyrifos/mouse, n=5), the animals were sacrificed. Blood samples and the tissues of interested were collected, weighted and counted. The percentage of injected does per gram (%ID/g) was calculated for each sample. The labeling yield of ¹³¹I-Chlorpyrifos was 93.5%, The radiochemical purity (RCP) was 96.9%. Biodistribution in mice demonstrated that¹³¹I-Chlorpyrifos was extensive, and the uptakes mainly occur in lung, stomach, small-intestine, colon, musle, and submaxillay gland, as indicated by their amount of 37.12%ID/g, 6.18%ID/g, 8.12%ID/g, 8.15%ID/g, 7.04%ID/g, and 7.02%ID/g at 10 min, respectively. And it was metabolized in liver and kidney, as indicated by their uptake of 4.34%ID/g and 8.50%ID/g at 5 min, and 0.22%ID/g and 0.69%ID/g at 4 h, respectively. In addition,¹³¹I-Chlorpyrifos was cleared out from blood quickly, and the uptake of¹³¹I-Chlorpyrifos in blood was 37.27%ID/g at 5 min, and decreased to 1.35%ID/g at 4 h post injection. In conclusion, ¹³¹I-Chlorpyrifos was stable in vitro and it was absorbed in lung and digestive tract, and it was metabolized mainly in liver and kidney, worthy of further investigation to trace the compound in vivo and in vitro.     

碘标记毒死蜱及其在小鼠体内的生物分布

为探讨¹³¹I标记毒死蜱（Chlorpyrifos, CPF）在小鼠体内的分布特点,采用Iodogen法对CPF进行¹³¹I标记,KM小鼠尾静脉注射¹³¹I-CPF（185 kBq/只,n=5）,分别于注射后5、10、30、60、120、240、1440 min取各脏器,计算每克组织摄取注射剂量的百分率（%ID/g）。结果显示,¹³¹I-CPF标记率达93.5%,放化纯度为96.9%,¹³¹I-CPF在小鼠体内广泛分布,主要经肺、胃、小肠、大肠、肌肉和颌下腺吸收,其放射性摄取率在注药后 10 min 时达高峰,分别为37.12%ID/g、6.18%ID/g、8.12%ID/g、8.15%ID/g、7.04%ID/g和7.02%ID/g;经肝和肾进行代谢,其放射性摄取率在5 min时分别为4.34%ID/g和8.50%ID/g, 4 h为0.22%ID/g和 0.69%ID/g。血液中放射性清除较快,放射性摄取率在注入后5 min时为37.27%ID/g,4 h为1.35%ID/g。碘标记CPF体外稳定,体内主要经肺和消化道吸收,肝、肾代谢,可用于进一步的微量示踪研究。     

Animal biodistribution, safety and validation study of dopamine transporter PET imaging agent ^18F-FECNT

This work was to investigate the pharmacologic characteristics of 18F-FECNT (2β-carbomethoxy-3β-(4-chlorophenyl)-8-(2-[18F]fluorcethyl)nortropane) as a dopamine transporter (DAT) PET imaging agent.Its partition coefficients were determined in n-octanol and phosphate buffer (PB) (pH 7.0 and pH 7.4).6-Hydroxydopamine (6-OHDA) left-sided lesioned Parkinsonian rats were established and validated by rotational behavior tests.Biodistribution in vivo in mice,autoradiography in normal and hemi-Parkinsonian rat brains,and toxicity test were performed.The results showed that partition coefficients were 34.14 (pH 7.0) and 56.41 (pH 7.4),respectively.Biodistribution exhibited rapid uptake and favorable retention in the mice brains.The major radioactivity was metabolized by the hepatic system.The autoradiography showed that 18F-FECNT was highly concentrated in striaturn,and that the left and the tight striatal uptake were symmetrical in normal SD rat brains.In left-sided lesioned PD rat brains,the striatal uptake of 18F-FECNT bilaterally decreased in comparison with normal rats.No significant uptake was visible in the 6-OHDA lesioned-sided striatal areas.The results demonstrated that 18F-FECNT binds to DAT was specific.Toxicity trial displayed that the acceptable dose per kilogram to mice was 625 times greater than that to human.These indicate that 18F-FECNT is a potentially safe and useful DAT PET imaging agent in the brain.     

红景天苷的碘标记及其在小鼠体内的分布

通过131碘标记红景天苷以探索红景天苷在神经母细胞(SH-SY5Y)中的摄取及在小鼠体内的代谢分布.采用氯胺-T法对红景天苷进行131碘标记;以聚酰胺薄膜为支持介质、V三氯甲烷:V甲醇:V丙酮:V水=6:3:1:1的下层液为展开剂,测定标记率及标记物放化纯;分析神经母细胞SH-SY5Y及肿瘤细胞MCF-7对131I-红景天苷的摄取;KM小鼠尾静脉注射131I-红景天苷(1.85 MBq/只,n=5),于5、10、30、60、120、240 min分别取心、肝、肺、肾、脾、肌、骨、脑、肠、血,称重、计数,计算每克组织百分注射剂量率(%ID/g).结果表明,131I-红景天苷标记率达98%,其放化纯在1、4、20 d分别为98.5%、97.3%、97.1%;SH-SY5Y对131I-红景天苷基本无摄取,在0.5-4 h内摄取维持在0.035%左右,而MCF-7则为0.1%;131I-红景天苷在体内主要通过肝代谢、肾排泄,其中肝和肾5 min%ID/g组织分别为7.71%和11.32%,4 h则分别下降为0.36%和0.3%;血液中清除也较快,5 min时为6.41%,4 h为0.35%;在脑中虽分布较少,但清除较慢,5 min时为0.27%,4 h为0.11%;在心、肺、脾、肌、骨及肠中分布不多.结论是,碘标红景天苷标记率高,标记物稳定;神经母细胞对131I-红景天苷基本无摄取.     

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.