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Dive into the research topics where Raffaele Ingenito is active.

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Featured researches published by Raffaele Ingenito.


FEBS Letters | 1998

Aggregates from mutant and wild‐type α‐synuclein proteins and NAC peptide induce apoptotic cell death in human neuroblastoma cells by formation of β‐sheet and amyloid‐like filaments

Omar M. A. El-Agnaf; Ross Jakes; Martin D. Curran; Derek Middleton; Raffaele Ingenito; Elisabetta Bianchi; Antonello Pessi; David Neill; Andrew Wallace

α‐Synuclein (α‐syn) protein and a fragment of it, called NAC, have been found in association with the pathological lesions of a number of neurodegenerative diseases. Recently, mutations in the α‐syn gene have been reported in families susceptible to an inherited form of Parkinsons disease. We have shown that human wild‐type α‐syn, mutant α‐syn(Ala30Pro) and mutant α‐syn(Ala53Thr) proteins can self‐aggregate and form amyloid‐like filaments. Here we report that aggregates of NAC and α‐syn proteins induced apoptotic cell death in human neuroblastoma SH‐SY5Y cells. These findings indicate that accumulation of α‐syn and its degradation products may play a major role in the development of the pathogenesis of these neurodegenerative diseases.


Bioorganic & Medicinal Chemistry | 2010

ApoA-I mimetic peptides promote pre-β HDL formation in vivo causing remodeling of HDL and triglyceride accumulation at higher dose.

Ester Carballo-Jane; Zhu Chen; Edward A. O’Neill; Jun Wang; Charlotte Burton; Ching H. Chang; Xun Chen; Suzanne S. Eveland; Betsy Frantz-Wattley; Karen Gagen; Brian K. Hubbard; Marina Ichetovkin; Silvi Luell; Roger Meurer; Xuelei Song; Alison M. Strack; Annunziata Langella; Simona Cianetti; Francesca Rech; Elena Capito; Simone Bufali; Maria Veneziano; Maria Verdirame; Fabio Bonelli; Edith Monteagudo; Antonello Pessi; Raffaele Ingenito; Elisabetta Bianchi

Reverse cholesterol transport promoted by HDL-apoA-I is an important mechanism of protection against atherosclerosis. We have previously identified apoA-I mimetic peptides by synthesizing analogs of the 22 amino acid apoA-I consensus sequence (apoA-I(cons)) containing non-natural aliphatic amino acids. Here we examined the effect of different aliphatic non-natural amino acids on the structure-activity relationship (SAR) of apoA-I mimetic peptides. These novel apoA-I mimetics, with long hydrocarbon chain (C(5-8)) amino acids incorporated in the amphipathic α helix of the apoA-I(cons), have the following properties: (i) they stimulate in vitro cholesterol efflux from macrophages via ABCA1; (ii) they associate with HDL and cause formation of pre-β HDL particles when incubated with human and mouse plasma; (iii) they associate with HDL and induce pre-β HDL formation in vivo, with a corresponding increase in ABCA1-dependent cholesterol efflux capacity ex vivo; (iv) at high dose they associate with VLDL and induce hypertriglyceridemia in mice. These results suggest our peptide design confers activities that are potentially anti-atherogenic. However a dosing regimen which maximizes their therapeutic properties while minimizing adverse effects needs to be established.


Bioorganic & Medicinal Chemistry Letters | 2010

Discovery and SAR of novel, potent and selective hexahydrobenzonaphthyridinone inhibitors of poly(ADP-ribose)polymerase-1 (PARP-1).

Caterina Torrisi; Monica Bisbocci; Raffaele Ingenito; Jesus M. Ontoria; Michael Rowley; Carsten Schultz-Fademrecht; Carlo Toniatti; Philip Jones

A novel hexahydrobenzonaphthyridinone PARP-1 pharmacophore is reported, subsequent SAR exploration around this scaffold led to selective PARP-1 inhibitors with low nanomolar enzyme potency, displaying good cellular activity and promising rat PK properties.


Archives of Biochemistry and Biophysics | 2017

Nuclear factor (erythroid-derived 2)-like 2 (NRF2) drug discovery: Biochemical toolbox to develop NRF2 activators by reversible binding of Kelch-like ECH-associated protein 1 (KEAP1)

Alberto Bresciani; Antonino Missineo; Mariana Gallo; Mauro Cerretani; Paola Fezzardi; Licia Tomei; Daniel O. Cicero; Sergio Altamura; Alessia Santoprete; Raffaele Ingenito; Elisabetta Bianchi; Robert Pacifici; Celia Dominguez; Ignacio Munoz-Sanjuan; Steven Harper; Leticia Toledo-Sherman; Larry Park

Mechanisms that activate innate antioxidant responses, as a way to mitigate oxidative stress at the site of action, hold much therapeutic potential in diseases, such as Parkinsons disease, Alzheimers disease and Huntingtons disease, where the use of antioxidants as monotherapy has not yielded positive results. The nuclear factor NRF2 is a transcription factor whose activity upregulates the expression of cell detoxifying enzymes in response to oxidative stress. NRF2 levels are modulated by KEAP1, a sensor of oxidative stress. KEAP1 binds NRF2 and facilitates its ubiquitination and subsequent degradation. Recently, compounds that reversibly disrupt the NRF2-KEAP1 interaction have been described, opening the field to a new era of safer NRF2 activators. This paper describes a set of new, robust and informative biochemical assays that enable the selection and optimization of non-covalent KEAP1 binders. These include a time-resolved fluorescence resonance energy transfer (TR-FRET) primary assay with high modularity and robustness, a surface plasmon resonance (SPR) based KEAP1 direct binding assay that enables the quantification and analysis of full kinetic binding parameters and finally a 1H-15N heteronuclear single quantum coherence (HSQC) NMR assay suited to study the interaction surface of KEAP1 with residue-specific information to validate the interaction of ligands in the KEAP1 binding site.


Journal of Pharmaceutical and Biomedical Analysis | 2018

A liquid chromatography high-resolution mass spectrometry in vitro assay to assess metabolism at the injection site of subcutaneously administered therapeutic peptides

Simone Esposito; Maria Lucia de Leonibus; Raffaele Ingenito; Elisabetta Bianchi; Laura Orsatti; Edith Monteagudo

Graphical abstract Figure. No caption available. HighlightsAn in vitro LC‐HRMS assay to predict peptide subcutaneous metabolism.in vitro half‐life is calculated and metabolites are identified in a single assay.Good in vitro/in vivo correlation was obtained for human and preclinical species.The assay can be used to improve peptide subcutaneous bioavailability. ABSTRACT Subcutaneous (SC) injection is the most common administration route for peptide therapeutics. Catabolism at the injection site can be a specific and major degradation pathway for many SC administered peptides. In some cases, it can significantly affect pharmacokinetics, particularly bioavailability, and have detrimental effects on the efficacy of the drug. This work describes a liquid chromatography‐high resolution mass spectrometry based in vitro assay to assess peptide metabolism in the SC tissue (SCiMetPep assay). The SCiMetPep assay was developed using human, Sprague‐Dawley rat and Göttingen minipig SC tissue homogenate supernatant, and allows for both determination of degradation rate (half‐life) of the parent peptide and identification of metabolites generated from enzymatic proteolysis. The assay was developed and validated using known peptides including human insulin and four GLP‐1 analogues (lixisenatide, exenatide, liraglutide and semaglutide). Different experimental parameters were evaluated in order to optimize the homogenization process of the SC tissue and the peptide incubation conditions. In vitro metabolism of these peptides was in good agreement with in vivo data reported in the literature. Finally, when SCiMetPep assay was applied on a series of structurally related peptides, a fairly good correlation was found between SC metabolic stability and bioavailability, suggesting that catabolism at the injection site can have a major role in the absorption, distribution, metabolism, and excretion (ADME) of peptide therapeutics. The SCiMetPep showed the ability to identify analogs with improved SC metabolic stability and hence higher bioavailability. The assay can be used in the early phases of drug discovery to identify peptide metabolic soft spots at the injection site and guide the peptide drug discovery process.


Archive | 2002

Structural studies of peptide inhibitors bound to hepatitis C virus protease yield insights into the mechanism of action of the enzyme

Antonello Pessi; Stefania Orrù; Paolo Ingallinella; Raffaele Ingenito; Uwe Koch; Piero Pucci; Elisabetta Bianchi

Much effort for a therapy against hepatitis C virus (HCV) is devoted to the search of inhibitors of the virally-encoded protease NS3, which is required for maturation of HCV polyprotein [1]. In order to cleave its substrates NS3 must form a complex with the 54residue viral cofactor protein NS4A, whose activity is mimicked by a synthetic peptide (Pep4A) corresponding to amino acids 21-34 [1]. Binding of Pep4A was shown to induce important tertiary structure changes in the enzyme [2]. We have recently developed substrate-derived hexapeptide inhibitors of NS3 with affinities in the low nanomolar range [3]. Structure-activity relationships, molecular modelling, site-directed mutagenesis and most recently NMR have been used to gain knowledge about the salient features of inhibitor binding [3-5]. In the present work we have further studied the interaction between NS3, NS4A and the inhibitors, and found that major conformational changes take place in the enzyme upon binary and ternary complex formation.


Archive | 2002

Engineering and chemical synthesis of the HCV protease transmembrane protein cofactor NS4A

Elisabetta Bianchi; Raffaele Ingenito; Reyna J. Simon; Antonello Pessi

In the course of our studies on the obligatory cofactor protein, NS4A, of the serine protease NS3 of Hepatitis C virus (HCV), we had to address two problems common to the study of transmembrane (TM) proteins: synthetic difficulties and poor solubility. NS3 is an HCV-encoded enzyme required for maturation of the viral polyprotein; to do so however, it must form a non-covalent complex [1] with the 54 residue protein NS4A (Table 1), which has been predicted to be a type I TM protein. So far any attempt at preparing NS4A by recDNA failed, and to date no protein is available for study.


Journal of the American Chemical Society | 1999

Solid Phase Synthesis of Peptide C-Terminal Thioesters by Fmoc/t-Bu Chemistry

Raffaele Ingenito; Elisabetta Bianchi; and Daniela Fattori; Antonello Pessi


Bioorganic & Medicinal Chemistry Letters | 2006

A series of novel, potent, and selective histone deacetylase inhibitors.

Philip Jones; Sergio Altamura; Prasun K. Chakravarty; Ottavia Cecchetti; Raffaele De Francesco; Paola Gallinari; Raffaele Ingenito; Peter T. Meinke; Alessia Petrocchi; Michael Rowley; Rita Scarpelli; Sergio Serafini; Christian Steinkühler


Biochemistry | 2000

Optimization of the P‘-Region of Peptide Inhibitors of Hepatitis C Virus NS3/4A Protease

Paolo Ingallinella; Elisabetta Bianchi; Raffaele Ingenito; Uwe Koch; Christian Steinkühler; Sergio Altamura; Antonello Pessi

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Antonello Pessi

École Normale Supérieure

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Rita Scarpelli

Istituto Italiano di Tecnologia

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