Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61

Andrea Calcaterra; Valentina Iovine; Bruno Botta; Deborah Quaglio; Ilaria D’Acquarica; Alessia Ciogli; Antonia Iazzetti; Romina Alfonsi; Ludovica Lospinoso Severini; Paola Infante; Lucia Di Marcotullio; Mattia Mori; Francesca Ghirga

doi:10.1080/14756366.2017.1419221

Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61

Andrea Calcaterra, Valentina Iovine, Bruno Botta, Deborah Quaglio, Ilaria D’Acquarica, Alessia Ciogli, Antonia Iazzetti, Romina Alfonsi, Ludovica Lospinoso Severini, Paola Infante, Lucia Di Marcotullio^*, Mattia Mori^*, Francesca Ghirga

^*Autore corrispondente per questo lavoro

Risultato della ricerca: Contributo in rivista › Articolo in rivista

Abstract

This work aims at elucidating the mechanism and kinetics of hydrolysis of GANT61, the first and most-widely used inhibitor of the Hedgehog (Hh) signalling pathway that targets Glioma-associated oncogene homologue (Gli) proteins, and at confirming the chemical nature of its bioactive form. GANT61 is poorly stable under physiological conditions and rapidly hydrolyses into an aldehyde species (GANT61-A), which is devoid of the biological activity against Hh signalling, and a diamine derivative (GANT61-D), which has shown inhibition of Gli-mediated transcription. Here, we combined chemical synthesis, NMR spectroscopy, analytical studies, molecular modelling and functional cell assays to characterise the GANT61 hydrolysis pathway. Our results show that GANT61-D is the bioactive form of GANT61 in NIH3T3 Shh-Light II cells and SuFu−/− mouse embryonic fibroblasts, and clarify the structural requirements for GANT61-D binding to Gli1. This study paves the way to the design of GANT61 derivatives with improved potency and chemical stability.

Lingua originale	English
pagine (da-a)	349-358
Numero di pagine	10
Rivista	Journal of Enzyme Inhibition and Medicinal Chemistry
Volume	33
DOI	https://doi.org/10.1080/14756366.2017.1419221
Stato di pubblicazione	Pubblicato - 2018

Keywords

Animals
Dose-Response Relationship, Drug
Fibroblasts
GANT61
Gli inhibitor
Hedgehog Proteins
Hedgehog pathway
Hydrolysis
Kinetics
Mice
Models, Molecular
Molecular Structure
NIH 3T3 Cells
Pyridines
Pyrimidines
Signal Transduction
Structure-Activity Relationship
bioactive form
chemical stability

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10.1080/14756366.2017.1419221

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Calcaterra, A., Iovine, V., Botta, B., Quaglio, D., D’Acquarica, I., Ciogli, A., Iazzetti, A., Alfonsi, R., Lospinoso Severini, L., Infante, P., Di Marcotullio, L., Mori, M., & Ghirga, F. (2018). Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61. Journal of Enzyme Inhibition and Medicinal Chemistry, 33, 349-358. https://doi.org/10.1080/14756366.2017.1419221

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title = "Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61",

abstract = "This work aims at elucidating the mechanism and kinetics of hydrolysis of GANT61, the first and most-widely used inhibitor of the Hedgehog (Hh) signalling pathway that targets Glioma-associated oncogene homologue (Gli) proteins, and at confirming the chemical nature of its bioactive form. GANT61 is poorly stable under physiological conditions and rapidly hydrolyses into an aldehyde species (GANT61-A), which is devoid of the biological activity against Hh signalling, and a diamine derivative (GANT61-D), which has shown inhibition of Gli-mediated transcription. Here, we combined chemical synthesis, NMR spectroscopy, analytical studies, molecular modelling and functional cell assays to characterise the GANT61 hydrolysis pathway. Our results show that GANT61-D is the bioactive form of GANT61 in NIH3T3 Shh-Light II cells and SuFu−/− mouse embryonic fibroblasts, and clarify the structural requirements for GANT61-D binding to Gli1. This study paves the way to the design of GANT61 derivatives with improved potency and chemical stability.",

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author = "Andrea Calcaterra and Valentina Iovine and Bruno Botta and Deborah Quaglio and Ilaria D{\textquoteright}Acquarica and Alessia Ciogli and Antonia Iazzetti and Romina Alfonsi and {Lospinoso Severini}, Ludovica and Paola Infante and {Di Marcotullio}, Lucia and Mattia Mori and Francesca Ghirga",

year = "2018",

doi = "10.1080/14756366.2017.1419221",

language = "English",

volume = "33",

pages = "349--358",

journal = "Journal of Enzyme Inhibition and Medicinal Chemistry",

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Calcaterra, A, Iovine, V, Botta, B, Quaglio, D, D’Acquarica, I, Ciogli, A, Iazzetti, A, Alfonsi, R, Lospinoso Severini, L, Infante, P, Di Marcotullio, L, Mori, M & Ghirga, F 2018, 'Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61', Journal of Enzyme Inhibition and Medicinal Chemistry, vol. 33, pagg. 349-358. https://doi.org/10.1080/14756366.2017.1419221

TY - JOUR

T1 - Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61

AU - Calcaterra, Andrea

AU - Iovine, Valentina

AU - Botta, Bruno

AU - Quaglio, Deborah

AU - D’Acquarica, Ilaria

AU - Ciogli, Alessia

AU - Iazzetti, Antonia

AU - Alfonsi, Romina

AU - Lospinoso Severini, Ludovica

AU - Infante, Paola

AU - Di Marcotullio, Lucia

AU - Mori, Mattia

AU - Ghirga, Francesca

PY - 2018

Y1 - 2018

N2 - This work aims at elucidating the mechanism and kinetics of hydrolysis of GANT61, the first and most-widely used inhibitor of the Hedgehog (Hh) signalling pathway that targets Glioma-associated oncogene homologue (Gli) proteins, and at confirming the chemical nature of its bioactive form. GANT61 is poorly stable under physiological conditions and rapidly hydrolyses into an aldehyde species (GANT61-A), which is devoid of the biological activity against Hh signalling, and a diamine derivative (GANT61-D), which has shown inhibition of Gli-mediated transcription. Here, we combined chemical synthesis, NMR spectroscopy, analytical studies, molecular modelling and functional cell assays to characterise the GANT61 hydrolysis pathway. Our results show that GANT61-D is the bioactive form of GANT61 in NIH3T3 Shh-Light II cells and SuFu−/− mouse embryonic fibroblasts, and clarify the structural requirements for GANT61-D binding to Gli1. This study paves the way to the design of GANT61 derivatives with improved potency and chemical stability.

AB - This work aims at elucidating the mechanism and kinetics of hydrolysis of GANT61, the first and most-widely used inhibitor of the Hedgehog (Hh) signalling pathway that targets Glioma-associated oncogene homologue (Gli) proteins, and at confirming the chemical nature of its bioactive form. GANT61 is poorly stable under physiological conditions and rapidly hydrolyses into an aldehyde species (GANT61-A), which is devoid of the biological activity against Hh signalling, and a diamine derivative (GANT61-D), which has shown inhibition of Gli-mediated transcription. Here, we combined chemical synthesis, NMR spectroscopy, analytical studies, molecular modelling and functional cell assays to characterise the GANT61 hydrolysis pathway. Our results show that GANT61-D is the bioactive form of GANT61 in NIH3T3 Shh-Light II cells and SuFu−/− mouse embryonic fibroblasts, and clarify the structural requirements for GANT61-D binding to Gli1. This study paves the way to the design of GANT61 derivatives with improved potency and chemical stability.

KW - Animals

KW - Dose-Response Relationship, Drug

KW - Fibroblasts

KW - GANT61

KW - Gli inhibitor

KW - Hedgehog Proteins

KW - Hedgehog pathway

KW - Hydrolysis

KW - Kinetics

KW - Mice

KW - Models, Molecular

KW - Molecular Structure

KW - NIH 3T3 Cells

KW - Pyridines

KW - Pyrimidines

KW - Signal Transduction

KW - Structure-Activity Relationship

KW - bioactive form

KW - chemical stability

KW - Animals

KW - Dose-Response Relationship, Drug

KW - Fibroblasts

KW - GANT61

KW - Gli inhibitor

KW - Hedgehog Proteins

KW - Hedgehog pathway

KW - Hydrolysis

KW - Kinetics

KW - Mice

KW - Models, Molecular

KW - Molecular Structure

KW - NIH 3T3 Cells

KW - Pyridines

KW - Pyrimidines

KW - Signal Transduction

KW - Structure-Activity Relationship

KW - bioactive form

KW - chemical stability

UR - http://hdl.handle.net/10807/200502

U2 - 10.1080/14756366.2017.1419221

DO - 10.1080/14756366.2017.1419221

M3 - Article

SN - 1475-6366

VL - 33

SP - 349

EP - 358

JO - Journal of Enzyme Inhibition and Medicinal Chemistry

JF - Journal of Enzyme Inhibition and Medicinal Chemistry

ER -

Chemical, computational and functional insights into the chemical stability of the Hedgehog pathway inhibitor GANT61

Abstract

Keywords

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