title: Combining X-ray crystallography and molecular modeling toward the optimization of pyrazolo[3,4-d]pyrimidines as potent c-Src inhibitors active in vivo against neuroblastoma.
abstract: c-Src is a tyrosine kinase belonging to the Src-family kinases. It is overexpressed and/or hyperactivated in a variety of cancer cells, thus its inhibition has been predicted to have therapeutic effects in solid tumors. Recently, the pyrazolo[3,4-d]pyrimidine 3 was reported as a dual c-Src/Abl inhibitor. Herein we describe a multidisciplinary drug discovery approach for the optimization of the lead 3 against c-Src. Starting from the X-ray crystal structure of c-Src in complex with 3, Monte Carlo free energy perturbation calculations were applied to guide the design of c-Src inhibitors with improved activities. As a result, the introduction of a meta hydroxyl group on the C4 anilino ring was computed to be particularly favorable. The potency of the synthesized inhibitors was increased with respect to the starting lead 3. The best identified compounds were also found active in the inhibition of neuroblastoma cell proliferation. Furthermore, compound 29 also showed in vivo activity in xenograft model using SH-SY5Y cells.
authors.affiliation: Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena Via Aldo Moro 2, 53100 Siena, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Tintori
authors.forename: Anna Lucia
authors.initials: AL
authors.lastname: Fallacara
authors.forename: Marco
authors.initials: M
authors.lastname: Radi
authors.forename: Claudio
authors.initials: C
authors.lastname: Zamperini
authors.forename: Elena
authors.initials: E
authors.lastname: Dreassi
authors.forename: Emmanuele
authors.initials: E
authors.lastname: Crespan
authors.forename: Giovanni
authors.initials: G
authors.lastname: Maga
authors.forename: Silvia
authors.initials: S
authors.lastname: Schenone
authors.forename: Francesca
authors.initials: F
authors.lastname: Musumeci
authors.forename: Chiara
authors.initials: C
authors.lastname: Brullo
authors.forename: André
authors.initials: A
authors.lastname: Richters
authors.forename: Francesca
authors.initials: F
authors.lastname: Gasparrini
authors.forename: Adriano
authors.initials: A
authors.lastname: Angelucci
authors.forename: Claudio
authors.initials: C
authors.lastname: Festuccia
authors.forename: Simona
authors.initials: S
authors.lastname: Delle Monache
authors.forename: Daniel
authors.initials: D
authors.lastname: Rauh
authors.forename: Maurizio
authors.initials: M
authors.lastname: Botta
mesh_terms: D000818:Animals; D000081247:CSK Tyrosine-Protein Kinase; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D015195:Drug Design; D055808:Drug Discovery; D006801:Humans; D008297:Male; D008819:Mice, Nude; D008956:Models, Chemical; D008958:Models, Molecular; D015394:Molecular Structure; D009010:Monte Carlo Method; D009447:Neuroblastoma / Q000188:drug therapy* / Q000473:pathology; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D011720:Pyrazoles / Q000737:chemistry / Q000494:pharmacology*; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D013816:Thermodynamics; D023041:Xenograft Model Antitumor Assays; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism
chemical_list: C000597230:N-(3-bromophenyl)-1-(2-chloro-2-phenylethyl)-6-((2-morpholin-4-ylethyl)thio)-1H-pyrazolo(3,4-d)pyrimidin-4-amine; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human