Gao Feng, Sihver Wiebke, Jurischka Christoph, Bergmann Ralf, Haase-Kohn Cathleen, Mosch Birgit, Steinbach Jörg, Carta Davide, Bolzati Cristina, Calderan Andrea, Pietzsch Jens, Pietzsch Hans-Jürgen
Helmholtz-Zentrum Dresden-Rossendorf, Institute of Radiopharmaceutical Cancer Research, Bautzner Landstraße 400, 01328, Dresden, Germany.
Department of Chemistry and Food Chemistry, Technische Universität Dresden, 01062, Dresden, Germany.
Amino Acids. 2016 Mar;48(3):833-847. doi: 10.1007/s00726-015-2131-x. Epub 2015 Dec 7.
The melanocortin-1 receptor (MC1R) plays an important role in melanoma growth, angiogenesis and metastasis, and is overexpressed in melanoma cells. α-Melanocyte stimulating hormone (α-MSH) and derivatives are known to bind with high affinity at this receptor that provides the potential for selective targeting of melanoma. In this study, one linear α-MSH-derived peptide Nle-Asp-His-D-Phe-Arg-Trp-Gly-NH2 (NAP-NS1) without linker and with εAhx-β-Ala linker, and a cyclic α-MSH derivative, [Lys-Glu-His-D-Phe-Arg-Trp-Glu]-Arg-Pro-Val-NH2 (NAP-NS2) with εAhx-β-Ala linker were conjugated with p-SCN-Bn-NOTA and labeled with (64)Cu. Radiochemical and radiopharmacological investigations were performed with regard to transchelation, stability, lipophilicity and in vitro binding assays as well as biodistribution in healthy rats. No transchelation reactions, but high metabolic stability and water solubility were demonstrated. The linear derivatives showed higher affinity than the cyclic one. [(64)Cu]Cu-NOTA-εAhx-β-Ala-NAP-NS1 ([(64)Cu]Cu-2) displayed rapid cellular association and dissociation in murine B16F10 cell homogenate. All [(64)Cu]Cu-labeled conjugates exhibited affinities in the low nanomolar range in B16F10. [(64)Cu]Cu-2 showed also high affinity in human MeWo and TXM13 cell homogenate. In vivo studies suggested that [(64)Cu]Cu-2 was stable, with about 85 % of intact peptide in rat plasma at 2 h p.i. Biodistribution confirmed the renal pathway as the major elimination route. The uptake of [(64)Cu]Cu-2 in the kidney was 5.9 % ID/g at 5 min p.i. and decreased to 2.0 % ID/g at 60 min p.i. Due to the prospective radiochemical and radiopharmacological properties of the linear α-MSH derivative [(64)Cu]Cu-2, this conjugate is a promising candidate for tracer development in human melanoma imaging.
黑皮质素-1受体(MC1R)在黑色素瘤的生长、血管生成和转移中起重要作用,且在黑色素瘤细胞中过表达。已知α-黑素细胞刺激素(α-MSH)及其衍生物能与该受体高亲和力结合,这为黑色素瘤的选择性靶向治疗提供了可能。在本研究中,一种无连接子的线性α-MSH衍生肽Nle-Asp-His-D-Phe-Arg-Trp-Gly-NH2(NAP-NS1)以及带有εAhx-β-Ala连接子的该肽,还有一种带有εAhx-β-Ala连接子的环状α-MSH衍生物[Lys-Glu-His-D-Phe-Arg-Trp-Glu]-Arg-Pro-Val-NH2(NAP-NS2)与p-SCN-Bn-NOTA偶联并标记上(64)Cu。针对转螯合、稳定性、亲脂性以及体外结合试验和在健康大鼠体内的生物分布进行了放射化学和放射药理学研究。结果表明不存在转螯合反应,但具有高代谢稳定性和水溶性。线性衍生物显示出比环状衍生物更高的亲和力。[(64)Cu]Cu-NOTA-εAhx-β-Ala-NAP-NS1([(64)Cu]Cu-2)在小鼠B16F10细胞匀浆中表现出快速的细胞结合和解离。所有[(64)Cu]Cu标记的偶联物在B16F10中表现出低纳摩尔范围内的亲和力。[(64)Cu]Cu-2在人MeWo和TXM13细胞匀浆中也显示出高亲和力。体内研究表明[(64)Cu]Cu-2是稳定的,在注射后2小时大鼠血浆中约85%为完整肽。生物分布证实肾脏途径是主要的清除途径。[(64)Cu]Cu-2在注射后5分钟时在肾脏中的摄取量为5.9% ID/g,在注射后60分钟时降至2.0% ID/g。由于线性α-MSH衍生物[(64)Cu]Cu-2具有预期的放射化学和放射药理学特性,该偶联物是用于人类黑色素瘤成像示踪剂开发的有前景的候选物。