title: Clickable, hydrophilic ligand for fac-[M(I)(CO)3](+) (M = Re/(99m)Tc) applied in an S-functionalized α-MSH peptide.
abstract: The copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) click reaction was used to incorporate alkyne-functionalized dipicolylamine (DPA) ligands (1 and 3) for fac-[M(I)(CO)3](+) (M = Re/(99m)Tc) complexation into an α-melanocyte stimulating hormone (α-MSH) peptide analogue. A novel DPA ligand with carboxylate substitutions on the pyridyl rings (3) was designed to increase the hydrophilicity and to decrease in vivo hepatobiliary retention of fac-[(99m)Tc(I)(CO)3](+) complexes used in single photon emission computed tomography (SPECT) imaging studies with targeting biomolecules. The fac-[Re(I)(CO)3(3)] complex (4) was used for chemical characterization and X-ray crystal analysis prior to radiolabeling studies between 3 and fac-[(99m)Tc(I)(OH2)3(CO)3](+). The corresponding (99m)Tc complex (4a) was obtained in high radiochemical yields, was stable in vitro for 24 h during amino acid challenge and serum stability assays, and showed increased hydrophilicity by log P analysis compared to an analogous complex with nonfunctionalized pyridine rings (2a). An α-MSH peptide functionalized with an azide was labeled with fac-[M(I)(CO)3](+) using both click, then chelate (CuAAC reaction with 1 or 3 followed by metal complexation) and chelate, then click (metal complexation of 1 and 3 followed by CuAAC with the peptide) strategies to assess the effects of CuAAC conditions on fac-[M(I)(CO)3](+) complexation within a peptide framework. The peptides from the click, then chelate strategy had different HPLC tR's and in vitro stabilities compared to those from the chelate, then click strategy, suggesting nonspecific coordination of fac-[M(I)(CO)3](+) using this synthetic route. The fac-[M(I)(CO)3](+)-complexed peptides from the chelate, then click strategy showed >90% stability during in vitro challenge conditions for 6 h, demonstrated high affinity and specificity for the melanocortin 1 receptor (MC1R) in IC50 analyses, and led to moderately high uptake in B16F10 melanoma cells. Log P analysis of the (99m)Tc-labeled peptides confirmed the enhanced hydrophilicity of the peptide bearing the novel, carboxylate-functionalized DPA chelate (10a') compared to the peptide with the unmodified DPA chelate (9a'). In vivo biodistribution analysis of 9a' and 10a' showed moderate tumor uptake in a B16F10 melanoma xenograft mouse model with enhanced renal uptake and surprising intestinal uptake for 10a' compared to predominantly hepatic accumulation for 9a'. These results, coupled with the versatility of CuAAC, suggests this novel, hydrophilic chelate can be incorporated into numerous biomolecules containing azides for generating targeted fac-[M(I)(CO)3](+) complexes in future studies.
authors.affiliation: Department of Chemistry, Washington State University , Pullman, Washington 99164, United States.
authors.forename: Benjamin B
authors.initials: BB
authors.lastname: Kasten
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Ma
authors.forename: Hongguang
authors.initials: H
authors.lastname: Liu
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Hayes
authors.forename: Charles L
authors.initials: CL
authors.lastname: Barnes
authors.forename: Shibo
authors.initials: S
authors.lastname: Qi
authors.forename: Kai
authors.initials: K
authors.lastname: Cheng
authors.forename: Shalina C
authors.initials: SC
authors.lastname: Bottorff
authors.forename: Winston S
authors.initials: WS
authors.lastname: Slocumb
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Cheng
authors.forename: Paul D
authors.initials: PD
authors.lastname: Benny
mesh_terms: D000588:Amines / Q000737:chemistry*; D000818:Animals; D002248:Carbon Monoxide / Q000737:chemistry*; D057930:Click Chemistry; D056831:Coordination Complexes / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics*; D005260:Female; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D008546:Melanoma, Experimental / Q000175:diagnosis*; D051379:Mice; D008810:Mice, Inbred C57BL; D010848:Picolinic Acids / Q000737:chemistry*; D019275:Radiopharmaceuticals / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics*; D012211:Rhenium / Q000737:chemistry*; D013667:Technetium / Q000737:chemistry*; D014018:Tissue Distribution; D015899:Tomography, Emission-Computed, Single-Photon; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000737:chemistry*
chemical_list: C501847:2,2'-dipicolylamine; D000588:Amines; D056831:Coordination Complexes; D008024:Ligands; D010848:Picolinic Acids; D019275:Radiopharmaceuticals; D000521:alpha-MSH; D012211:Rhenium; D013667:Technetium; D002248:Carbon Monoxide