Design of highly specific ligands of fibrin for therapeutic applications.
The work presented here is aimed at designing high-affinity ligands for the fibrin gamma (312-324) epitope. This epitope is specific for fibrin recognition, as it is exposed only on the fibrin surface, while in fibrinogen it is buried in the protein bulk. This property makes it very useful for therapeutic applications. In fact, it may be exploited in driving systems for targeted delivery of thrombolytic drugs toward the specific compartment where they are needed. It will then allow avoidance of serious unwanted side effects produced by a conventional systemic administration. The approach chosen for designing putative ligands is based on the known three-dimensional (3D) structure of the epitope. A wide virtual library made up of oligo-peptides and analogues designed by a combinatorial approach, on the basis of chemical complementarity criteria, has been screened by means of a docking/scoring approach (DOCK program). The peculiarity of the problem under study required a considerable effort in finding a method enabling the experimental validation of the design work results. In fact the selected biological target is absolutely new, so that neither a endogenous, nor a synthetic high-affinity ligand is known up to now. It does not allow for the validation of computational results by means of classical binding tests based on the use of known labeled high-affinity ligands. Preliminary binding essayes were so carried out by means of the Plasmon Surface Resonance (PSR) technique. The experimental results suggested that most of the molecules predicted to be good ligands by means of the selected computational tools, could carry the wanted affinity toward the selected target.
Bianucci
Anna Maria
AM
Department of Pharmaceutical Science, University of Pisa, via Bonanno Pisano 6, 56126 Pisa, Italy.
Massarelli
Ilaria
I
Chiellini
Federica
F
Eidelman
Chaim
C
Chiellini
Emo
E
D000595:Amino Acid Sequence; D000818:Animals; D001672:Biocompatible Materials / Q000138:chemical synthesis / Q000737:chemistry; D002417:Cattle; D015195:Drug Design; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism; D005337:Fibrin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008422:Materials Testing; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013816:Thermodynamics
D001672:Biocompatible Materials; D000939:Epitopes; D005353:Fibronectins; D008024:Ligands; D009842:Oligopeptides; D019151:Peptide Library; D005337:Fibrin