title: Branched porphyrins as functional scaffolds for multisite bioconjugation.
abstract: Bioconjugation is a rapidly expanding field because of the numerous potential applications of bioconjugate materials. We explored the usefulness of branched porphyrins as rigid scaffolds, bearing multiple sites for bioconjugation. To this end, we first selected the tetrakis(p-[aminomethyl] phenyl) porphyrin (TAMPP) macrocycle and developed a straightforward synthetic protocol, able to provide the desired tetraphenylporphyrin, carrying four functional amino groups. The partially protection of the amino groups by tert-butoxy-carbonyl allowed the selective and specific decoration of the porphyrin with different peptide sequences. To explore the utility of the macrocycle as molecular scaffold for bioconjugation, we selected peptide sequences able to function as thrombin inhibitors. In particular, two peptide sequences, named CS3 and ES7, able to interact, respectively, with the thrombin catalytic site and the fibrinogen recognition exosite, were joined onto the porphyrin macrocycle, providing the multisite-directed inhibitor CS3-TAMPP-ES7. This multisite inhibitor and its Mn(III) complex are able to inhibit α-thrombin-catalyzed hydrolysis of Tos-Gly-Pro-Arg-nitroanilide with inhibition constants in the micromolar range, as well as the hydrolysis of the natural substrate fibrinogen. The inhibitor is resistant against enzymatic degradation by thrombin and is highly selective. The Mn(III) complex is capable of interacting with clot-bound thrombin and partially inhibits clot growth in the presence of fibrinogen. The results herein reported are very promising, suggesting the potential of the newly developed conjugate as new imaging agents for clot detection.
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Mireille
authors.initials: M
authors.lastname: Engelen
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Lombardi
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Rosa
authors.initials: R
authors.lastname: Vitale
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Liliana
authors.initials: L
authors.lastname: Lista
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.|IBB, CNR, Napoli, Italy.
authors.forename: Ornella
authors.initials: O
authors.lastname: Maglio
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Pavone
authors.affiliation: Department of Chemical Sciences, University of Naples "Federico II," Complesso Universitario Monte S. Angelo, Napoli, Italy.
authors.forename: Flavia
authors.initials: F
authors.lastname: Nastri
mesh_terms: D000991:Antithrombins / Q000737:chemistry / Q000494:pharmacology; D001672:Biocompatible Materials / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D008956:Models, Chemical; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry*; D011166:Porphyrins / Q000737:chemistry*; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000991:Antithrombins; D001672:Biocompatible Materials; D010455:Peptides; D011166:Porphyrins; D013917:Thrombin
keywords: metalloporphyrins; peptide-porphyrin conjugates; porphyrin scaffold; porphyrin synthesis; thrombin inhibitors