title: Synthesis and in Vitro and in Vivo Evaluation of MMP-12 Selective Optical Probes.
abstract: In designing new tracers consisting of a small peptide conjugated to a reporter of comparable size, particular attention needs to be paid to the selection of the reporter group, which can dictate both the in vitro and the in vivo performances of the whole conjugate. In the case of fluorescent tracers, this is particularly true given the large numbers of available dye moieties differing in their structures and properties. Here, we have investigated the in vitro and in vivo properties of a novel series of MMP-12 selective probes composed of cyanine dyes varying in their structure, net charge, and hydrophilic character, tethered through a linker to a potent and specific MMP-12 phosphinic pseudopeptide inhibitor. The impact of linker length has been also explored. The crystallographic structure of one tracer in complex with MMP-12 has been obtained, providing the first crystal structure of a Cy5.5-derived probe and confirming that the binding of the targeting moiety is unaffected. MMP-12 remains the tracers' privileged target, as attested by their affinity selectivity profile evaluated in solution toward a panel of 12 metalloproteases. In vivo assessment of four selected probes has highlighted not only the impact of the dye structure but also that of the linker length on the probes' blood clearance rates and their biodistributions. These experiments have also provided valuable data on the stability of the dye moieties in vivo. This has permitted the identification of one probe, which combines favorable binding to MMP-12 in solution and on cells with optimized in vivo performance including blood clearance rate suitable for short-time imaging. Through this series of tracers, we have identified various critical factors modulating the tracers' in vivo behavior, which is both useful for the development and optimization of MMP-12 selective radiolabeled tracers and informative for the design of fluorescent probes in general.
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Thomas
authors: T
authors: Bordenave
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Marion
authors: M
authors: Helle
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Fabrice
authors: F
authors: Beau
authors: Department of Chemistry, Laboratory of Organic Chemistry, University of Athens , Panepistimiopolis, Zografou, Athens 15771, Greece.
authors: Dimitris
authors: D
authors: Georgiadis
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Livia
authors: L
authors: Tepshi
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Mylène
authors: M
authors: Bernes
authors: Cardiovascular Molecular Imaging Laboratory, Section of Cardiovascular Medicine and Yale Cardiovascular Research Center, Yale University School of Medicine , New Haven, Connecticut 06511, United States.|Veterans Affairs Connecticut Healthcare System , West Haven, Connecticut 06516, United States.
authors: Yunpeng
authors: Y
authors: Ye
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Laure
authors: L
authors: Levenez
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Enora
authors: E
authors: Poquet
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Hervé
authors: H
authors: Nozach
authors: Cardiovascular Molecular Imaging Laboratory, Section of Cardiovascular Medicine and Yale Cardiovascular Research Center, Yale University School of Medicine , New Haven, Connecticut 06511, United States.|Veterans Affairs Connecticut Healthcare System , West Haven, Connecticut 06516, United States.
authors: Mahmoud
authors: M
authors: Razavian
authors: Cardiovascular Molecular Imaging Laboratory, Section of Cardiovascular Medicine and Yale Cardiovascular Research Center, Yale University School of Medicine , New Haven, Connecticut 06511, United States.|Veterans Affairs Connecticut Healthcare System , West Haven, Connecticut 06516, United States.
authors: Jakub
authors: J
authors: Toczek
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Enrico A
authors: EA
authors: Stura
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Vincent
authors: V
authors: Dive
authors: Cardiovascular Molecular Imaging Laboratory, Section of Cardiovascular Medicine and Yale Cardiovascular Research Center, Yale University School of Medicine , New Haven, Connecticut 06511, United States.|Veterans Affairs Connecticut Healthcare System , West Haven, Connecticut 06516, United States.
authors: Mehran M
authors: MM
authors: Sadeghi
authors: Service d'ingénierie moléculaire des protéines (SIMOPRO), IBITECS, CEA, Université Paris-Saclay , Gif-sur-Yvette F-91191, France.
authors: Laurent
authors: L
authors: Devel
mesh_terms: D000818:Animals; D002232:Carbocyanines; D060326:Chemistry Techniques, Synthetic; D018360:Crystallography, X-Ray; D006367:HeLa Cells; D006801:Humans; D053508:Matrix Metalloproteinase 12 / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D008810:Mice, Inbred C57BL; D057054:Molecular Imaging / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism* / Q000493:pharmacokinetics; D055095:Optics and Photonics / Q000379:methods; D010455:Peptides / Q000737:chemistry; D014018:Tissue Distribution
chemical_list: C098793:CY5.5 cyanine dye; D002232:Carbocyanines; D015335:Molecular Probes; D010455:Peptides; C557203:RXP470.1 peptide; D053508:Matrix Metalloproteinase 12