title: A cascade screening approach for the identification of Bcr-Abl myristate pocket binders active against wild type and T315I mutant.
abstract: The major clinical challenge in drug-resistant chronic myelogenous leukemia (CML) is currently represented by the Bcr-Abl T315I mutant, which is unresponsive to treatment with common first and second generation ATP-competitive tyrosine kinase inhibitors (TKIs). Allosteric inhibition of Bcr-Abl represent a new frontier in the fight against resistant leukemia and few candidates have been identified in the last few years. Among these, myristate pocket (MP) binders discovered by Novartis (e.g. GNF2/5) showed promising results, although they proved to be active against the T315I mutant only in combination with first and second generation ATP-competitive inhibitors. Here we used a cascade screening approach based on sequential fluorescence polarization (FP) screening, in silico docking/dynamics studies and kinetic-enzymatic studies to identify novel MP binders. A pyrazolo[3,4-d]pyrimidine derivative (6) has been identified as a promising allosteric inhibitor active on 32D leukemia cell lines (expressing Bcr-Abl WT and T315I) with no need of combination with any ATP-competitive inhibitor.
authors.affiliation: Dipartimento Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via A. De Gasperi 2, I-53100 Siena, Italy; P4T Group, Dipartimento di Farmacia, Università degli Studi di Parma, Viale delle Scienze, 27/A, 43124 Parma, Italy. Electronic address: marco.radi@unipr.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Radi
authors.affiliation: Department of Chemistry and Chemical Biology, Technical University of Dortmund, Otto-Hahn-Strasse 6, 44227 Dortmund, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Schneider
authors.affiliation: Dipartimento Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via A. De Gasperi 2, I-53100 Siena, Italy.
authors.forename: Anna Lucia
authors.initials: AL
authors.lastname: Fallacara
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", Via D. Montesano 49, 80131 Naples, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Botta
authors.affiliation: Istituto di Genetica Molecolare, IGM-CNR, Via Abbiategrasso 207, 27100 Pavia, Italy.
authors.forename: Emmanuele
authors.initials: E
authors.lastname: Crespan
authors.affiliation: Dipartimento Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, Via A. De Gasperi 2, I-53100 Siena, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Tintori
authors.affiliation: Istituto di Genetica Molecolare, IGM-CNR, Via Abbiategrasso 207, 27100 Pavia, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Maga
authors.affiliation: Istituto di Genetica Molecolare, IGM-CNR, Via Abbiategrasso 207, 27100 Pavia, Italy.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Kissova
authors.affiliation: Dipartimento di Scienze Cliniche Applicate e Biotecnologiche, Università degli Studi dell'Aquila, Via Vetoio, 67100 Coppito, L'Aquila, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Calgani
authors.affiliation: Department of Chemistry and Chemical Biology, Technical University of Dortmund, Otto-Hahn-Strasse 6, 44227 Dortmund, Germany.
authors.forename: André
authors.initials: A
authors.lastname: Richters
authors.affiliation: Dipartimento di Farmacia, Università degli Studi di Genova, Viale Benedetto XV, 3, 16132 Genova, Italy.
authors.forename: Franesca
authors.initials: F
authors.lastname: Musumeci
authors.affiliation: Department of Chemistry and Chemical Biology, Technical University of Dortmund, Otto-Hahn-Strasse 6, 44227 Dortmund, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Rauh
authors.affiliation: Dipartimento di Farmacia, Università degli Studi di Genova, Viale Benedetto XV, 3, 16132 Genova, Italy. Electronic address: schensil@unige.it.
authors.forename: Silvia
authors.initials: S
authors.lastname: Schenone
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy* / Q000473:pathology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D009226:Myristates / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D008565:Membrane Proteins; D009226:Myristates; D009363:Neoplasm Proteins; C546577:T351 protein, human
keywords: Allosteric; Bcr-Abl; Kinase; Leukemia; Myristate; Resistance; T315I