Tramarin, Anna <1990>
Development of multimethodological strategies for monitoring biorecognition phenomena of pharmaceutically relevant targets [Tesi di dottorato]
Alma Mater Studiorum - Università di Bologna, 2019-03-28

The comprehension of biorecognition phenomena is pivotal to clarify pathophysiological mechanisms and rationally develop new and more effective drugs. Amongst available analytical approaches to profile the binding partners, mass spectrometry (MS), circular dichroism (CD) and surface plasmon resonance (SPR) spectroscopies have been used to investigate two pharmaceutically relevant targets: human serum albumin (HSA) and human cholinesterases (ChEs). Because of the role of HSA in several biological functions and its use in clinical practice as biological drug, it represents an important object of research. In this scenario, two studies were conducted. In the first, the binding capacity of pharmaceutical-grade HSA for infusion was studied by implementing a CD spectroscopy-based assay. A clear impairment of the binding capacity caused by the stabilizers sodium octanoate and N-acetyltryptophan, along with the impossibility of removing octanoate by common approaches, were highlighted. In the second study, SPR- and affinity chromatography-MS-based assays enabled an initial characterization of the interaction between glycated HSA and the receptor for advanced glycation end products (RAGE), providing further insights into the biorecognition phenomenon and laying the groundwork for subsequent studies on AGEs as more representative players in diabetes. On the other hand, ChE enzymes are widely studied molecular targets in drug discovery for Alzheimer’s disease. Their inhibition still represents the main strategy to temporally counteract cognitive impairment and ameliorate patients’ quality of life. In this context, in solution functional assays were used for the identification of new potential inhibitors while a tailored SPR-based assay was developed to complement classic inhibition studies, helping the prioritization of favorite chemical scaffolds by providing affinity and kinetic parameters, including residence time. Overall, the tailored analytical strategies herein reported allowed to elucidate key biorecognition events, providing pivotal information which can help understanding pathological events as well as favoring the drug discovery process.

diritti: info:eu-repo/semantics/openAccess
In relazione con
Bartolini, Manuela
CHIM/08 - Chimica farmaceutica

Tesi di dottorato. | Lingua: en. | Paese: | BID: TD20009793