HIGH RESOLUTION MASS SPECTROMETRIC STRATEGIES IN DRUG DISCOVERY FOR THE INVESTIGATION OF COVALENT AND NON-COVALENT INTERACTIONS
Tesi di Dottorato
Data di Pubblicazione:
2014
Citazione:
HIGH RESOLUTION MASS SPECTROMETRIC STRATEGIES IN DRUG DISCOVERY FOR THE INVESTIGATION OF COVALENT AND NON-COVALENT INTERACTIONS / D. De Maddis ; tutor: G. Aldini; supervisore: M. Carini; coordinatore: E. Valoti. DIPARTIMENTO DI SCIENZE FARMACEUTICHE, 2014 Feb 25. 26. ciclo, Anno Accademico 2013. [10.13130/de-maddis-danilo_phd2014-02-25].
Abstract:
High resolution mass spectrometric strategies in drug discovery for the investigation of covalent and non covalent interactions
Dott. Danilo De Maddis
PhD tutor: Prof. Giancarlo Aldini
The research work here described was focused on the set-up and application of analytical methods for studying compounds effective as inhibitors of the advanced glycation end-products (AGEs) and advanced lipoxidation end-products (ALEs) as well as as antagonists of the receptors of AGEs (RAGE).
AGEs and ALEs represent a quite complex and heterogeneous class of compounds that are formed by different mechanisms, by heterogeneous precursors and can be formed either exogenously or endogenously.
AGEs represent a class of covalently modified proteins generated by oxidative and non-oxidative pathways, involving sugars or their degradation products. The term ALEs includes a variety of covalent adducts which are generated by the non-enzymatic reaction of reactive carbonyl species (RCS), produced by lipid peroxidation and lipid metabolism, with the nucleophilic residues of macromolecules, especially proteins.
AGEs and ALEs share some common properties, for example, both consist of non-enzymatic,
covalently modified proteins and oxidative stress is often (although not always) involved in the mechanism of their formation. Moreover some AGEs and ALEs have the same structure, since they arise from common precursors, as in the case of carboxymethyllysine (CML) which is generated by glyoxal, which in turn is formed by both lipid and sugar oxidative degradation
pathways [1].
Besides being considered as reliable biomarkers of oxidative damage, as well as predictors and prognostic factors, more recently, AGEs and ALEs have also been recognized as important pathogenetic factors of some oxidative based diseases, as supported by the following facts: 1) a strict correlation between the amount of AGEs/ALEs in tissues and fluids and disease states has been found, in both animal and human subjects; 2) a substantial amount of literature is now available reporting the molecular and cellular pathogenic mechanisms for the AGEs/ALEs involvement in the onset and progression of different oxidative-based diseases including diabetes [2], chronic renal failure [3], cardiovascular diseases [4] and neurological disorders [5]. The AGEs/ALEs damaging effect is mediated by different mechanisms, including the dysfunction of the proteins undergoing the oxidative modification, protein polymerization, signal transduction, immunoresponse and RAGE activation. Some of the biological effects are due to the loss of function of the target proteins undergoing the covalent modification, such as in the case of extracellular matrix proteins that lose their elastic and mechanical functions when modified as AGEs/ALEs and in particular, when cross-links are involved [6]. Other examples of a direct damaging effect of AGEs/ALEs can be ascribed to the covalent modification of enzymes and receptors that lose their activity due to the covalent modification involving the catalytic or binding site, or following a conformational change of the protein structure. Moreover AGEs and ALEs can be immunogenic.
Hence AGEs/ALEs are now considered as promising drug targets and a substantial effort is dedicated to delve the molecular strategies aimed at preventing, reducing or removing these protein oxidation products.
The different molecular approaches thus far reported can be grouped by considering at which level of the damaging AGEs/ALEs cascade they are effective and in particular if they act by inhibiting the AGEs/ALEs formation, accelerating their catabolism or blocking their biological effects.
The first level of action, the inhibition of AGEs/ALEs formation, also consists of different approac
Tipologia IRIS:
Tesi di dottorato
Keywords:
high ; resolution ; mass ; spectrometric ; drug ; investigation ; covalent ; interactions ; RAGE ; Orbitrap ; HPLC
Elenco autori:
D. DE MADDIS
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