Radiochemical investigations of (99m)Tc-N(3)S-X-BBN[7-14]NH(2): an in vitro/in vivo structure-activity relationship study where X = 0-, 3-, 5-, 8-, and 11-carbon tethering moieties.
Radiochemical investigations of (99m)Tc-N(3)S-X-BBN[7-14]NH(2): an in vitro/in vivo structure-activity relationship study where X = 0-, 3-, 5-, 8-, and 11-carbon tethering moieties.
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DOI：
10.1021/bc020034r
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发表时间：
2003
影响因子：
4.7
通讯作者：
C. Smith;H. Gali;G. Sieckman;C. Higginbotham;W. Volkert;T. Hoffman
中科院分区：
文献类型：
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作者：
C. Smith;H. Gali;G. Sieckman;C. Higginbotham;W. Volkert;T. Hoffman
蛙皮素（BBN）是一种由14个氨基酸组成的肽，是人胃泌素释放肽（GRP）的类似物，其以高亲和力和特异性结合GRP受体（GRPr）。GRPr在多种人类癌细胞上过表达，包括前列腺癌、乳腺癌、肺癌和胰腺癌。本研究的具体目的是开发（99 m）Tc-放射性标记的BBN类似物，在体内保持高特异性的GRPr。设计了通过固相肽合成（SPPS）的预选合成序列，以产生具有以下一般结构的N（3）S-BBN（N（3）S =二甲基甘氨酰-1-丝氨酰-1-半胱氨酰甘氨酰胺）缀合物：DMG-S-C-G-X-Q-W-A-V-G-H-L-M-（NH（2）），其中间隔基团X = 0（无间隔基）、ω-NH（2）（CH（2））（2）COOH、ω-NH（2）（CH（2））（4）COOH、ω-NH（2）（CH（2））（7）COOH或ω-NH（2）-（CH（2））（10）COOH。通过反相HPLC（RP-HPLC）纯化新的BBN构建体。电喷雾质谱（ES-MS）用于表征非金属化的BBN缀合物。通过葡萄糖酸Re（V）与N（3）S-X-BBN[7-14]NH（2）（X = 0个碳，β-Ala（β-丙氨酸），5-Ava（5-氨基戊酸），8-Aoc（8-氨基辛酸）和11-Aun（11-氨基十一烷酸））在温和加热下反应制备Re（V）-BBN缀合物。通过[Re（V）二甲基甘氨酰-1-丝氨酰-1-半胱氨酰甘氨酰胺]与5-Ava-BBN [7 - 14] NH（2）的反应也制备了Re-N（3）S-5-Ava-BBN [7 - 14] NH（2）。ES-MS用于确定新Re（V）缀合物的分子组成。通过将Na[（99 m）TcO（4）]与过量的SnCl（2）、葡萄糖酸钠和相应的配体反应，分别采用预标记、后标记和预标记、后标记的方法，在示踪剂水平制备了（99 m）Tc缀合物。在相同的反相高效液相色谱条件下，（99 m）Tc和Re（V）结合物的行为相似。体外和体内模型证明了新缀合物的生物完整性。
Bombesin (BBN), a 14 amino acid peptide, is an analogue of human gastrin releasing peptide (GRP) that binds to GRP receptors (GRPr) with high affinity and specificity. The GRPr is overexpressed on a variety of human cancer cells, including prostate, breast, lung, and pancreatic cancers. The specific aim of this study was to develop (99m)Tc-radiolabeled BBN analogues that maintain high specificity for the GRPr in vivo. A preselected synthetic sequence via solid-phase peptide synthesis (SPPS) was designed to produce N(3)S-BBN (N(3)S = dimethylglycyl-l-seryl-l-cysteinylglycinamide) conjugates with the following general structure: DMG-S-C-G-X-Q-W-A-V-G-H-L-M-(NH(2)), where the spacer group, X = 0 (no spacer), omega-NH(2)(CH(2))(2)COOH, omega-NH(2)(CH(2))(4)COOH, omega-NH(2)(CH(2))(7)COOH, or omega-NH(2)-(CH(2))(10)COOH. The new BBN constructs were purified by reversed phase-HPLC (RP-HPLC). Electrospray mass spectrometry (ES-MS) was used to characterize the nonmetalated BBN conjugates. Re(V)-BBN conjugates were prepared by the reaction of Re(V)gluconate with N(3)S-X-BBN[7-14]NH(2) (X = 0 carbons, beta-Ala (beta-alanine), 5-Ava (5-aminovaleric acid), 8-Aoc (8-aminooctanoic acid), and 11-Aun (11-aminoundecanoic acid)) with gentle heating. Re-N(3)S-5-Ava-BBN[7-14]NH(2) was also prepared by the reaction of [Re(V)dimethylglycyl-l-seryl-l-cysteinylglycinamide] with 5-Ava-BBN[7-14]NH(2). ES-MS was used to determine the molecular constitution of the new Re(V) conjugates. The (99m)Tc conjugates were prepared at the tracer level by each the prelabeling, post-conjugation and pre-conjugation, postlabeling approaches from the reaction of Na[(99m)TcO(4)] with excess SnCl(2), sodium gluconate, and corresponding ligand. The (99m)Tc and Re(V) conjugates behaved similarly under identical RP-HPLC conditions. In vitro and in vivo models demonstrated biological integrity of the new conjugates.