TY - JOUR
T1 - Structural Basis of Colchicine-Site targeting Acylhydrazones active against Multidrug-Resistant Acute Lymphoblastic Leukemia
AU - Cury, Nathália Moreno
AU - Mühlethaler, Tobias
AU - Laranjeira, Angelo Brunelli Albertoni
AU - Canevarolo, Rafael Renatino
AU - Zenatti, Priscila Pini
AU - Lucena-Agell, Daniel
AU - Barasoain, Isabel
AU - Song, Chunhua
AU - Sun, Dongxiao
AU - Dovat, Sinisa
AU - Yunes, Rosendo Augusto
AU - Prota, Andrea Enrico
AU - Steinmetz, Michel Olivier
AU - Díaz, José Fernando
AU - Yunes, José Andrés
N1 - Funding Information:
We thank Ganadería Fernando Díaz for calf brains for tubulin purification. The authors acknowledge networking contribution by the COST Action CM1407 “Challenging organic syntheses inspired by nature - from natural products chemistry to drug discovery.” J.F.D. is a member of the CIB Intramural Program “Molecular Machines for Better Life” (MACBET). N.M.C. was supported by a fellowship from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, 14/08247-8, and 17/14737-6). J.A.Y. received a Productivity fellowship from the Brazilian National Counsel of Technological and Scientific Development (CNPq 305896/2013-0 and 301596/2017-4). This work was supported in part by grants BFU2016-75319-R (AEI/FEDER, UE) (J.F.D.) from Ministerio de Economía y Competitividad. The crystal structure work was supported by grants from the Swiss National Science Foundation (31003A_166608, to M.O.S.) and by the COST action CM1407 (to M.O.S.). Part of the in vivo work was supported by R01CA209829 and R01CA213912, Hyundai Hope On Wheels Scholar Grant, Bear Necessities Pediatric Cancer Foundation, Alex's Lemonade Stand Foundation, the Four Diamonds Fund of the Pennsylvania State University College of Medicine, and the John Wawrynovic Leukemia Research Scholar Endowment (to S.D.). N.M.C. performed the majority of the experiments, contributed to the analyses and interpretation of the results, and drafted the manuscript. T.M. performed the crystal structure work and wrote the manuscript. A.B.A.L. performed the in vivo anti-leukemia experiment. R.R.C. supervised the project and contributed to the interpretation of the results. P.P.Z. performed the in vivo toxicity experiment. D.L.-A. contributed to the standardization and interpretation of biochemical results. I.B. contributed to the standardization and analyses of immunofluorescence assay. C.S. and S.D. supervised the in vivo assays with NALM6 and N6/ADR cell lines. D.S. performed mass spectrometry analyzes. R.A.Y. designed and provided the compounds used in the present work. A.E.P. and M.O.S. supervised the crystal structure work and critically reviewed the manuscript. J.F.D. supervised the biochemical work, contributed to the interpretation of the results, and critically reviewed the manuscript. J.A.Y. was responsible for the conception and design of study. He supervised the entire project and wrote the manuscript. Compounds 7, 12, and 21 are patented under WO 2013/075199 A1.
Funding Information:
N.M.C. was supported by a fellowship from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, 14/08247-8 , and 17/14737-6 ). J.A.Y. received a Productivity fellowship from the Brazilian National Counsel of Technological and Scientific Development ( CNPq 305896/2013-0 and 301596/2017-4 ). This work was supported in part by grants BFU2016-75319-R (AEI/FEDER, UE) (J.F.D.) from Ministerio de Economía y Competitividad . The crystal structure work was supported by grants from the Swiss National Science Foundation ( 31003A_166608 , to M.O.S.) and by the COST action CM1407 (to M.O.S.). Part of the in vivo work was supported by R01CA209829 and R01CA213912 , Hyundai Hope On Wheels Scholar Grant, Bear Necessities Pediatric Cancer Foundation , Alex’s Lemonade Stand Foundation , the Four Diamonds Fund of the Pennsylvania State University College of Medicine , and the John Wawrynovic Leukemia Research Scholar Endowment (to S.D.).
Publisher Copyright:
© 2019 The Authors
PY - 2019/11/22
Y1 - 2019/11/22
N2 - Tubulin is one of the best validated anti-cancer targets, but most anti-tubulin agents have unfavorable therapeutic indexes. Here, we characterized the tubulin-binding activity, the mechanism of action, and the in vivo anti-leukemia efficacy of three 3,4,5-trimethoxy-N-acylhydrazones. We show that all compounds target the colchicine-binding site of tubulin and that none is a substrate of ABC transporters. The crystal structure of the tubulin-bound N-(1′-naphthyl)-3,4,5-trimethoxybenzohydrazide (12) revealed steric hindrance on the T7 loop movement of β-tubulin, thereby rendering tubulin assembly incompetent. Using dose escalation and short-term repeated dose studies, we further report that this compound class is well tolerated to >100 mg/kg in mice. We finally observed that intraperitoneally administered compound 12 significantly prolonged the overall survival of mice transplanted with both sensitive and multidrug-resistant acute lymphoblastic leukemia (ALL) cells. Taken together, this work describes promising colchicine-site-targeting tubulin inhibitors featuring favorable therapeutic effects against ALL and multidrug-resistant cells.
AB - Tubulin is one of the best validated anti-cancer targets, but most anti-tubulin agents have unfavorable therapeutic indexes. Here, we characterized the tubulin-binding activity, the mechanism of action, and the in vivo anti-leukemia efficacy of three 3,4,5-trimethoxy-N-acylhydrazones. We show that all compounds target the colchicine-binding site of tubulin and that none is a substrate of ABC transporters. The crystal structure of the tubulin-bound N-(1′-naphthyl)-3,4,5-trimethoxybenzohydrazide (12) revealed steric hindrance on the T7 loop movement of β-tubulin, thereby rendering tubulin assembly incompetent. Using dose escalation and short-term repeated dose studies, we further report that this compound class is well tolerated to >100 mg/kg in mice. We finally observed that intraperitoneally administered compound 12 significantly prolonged the overall survival of mice transplanted with both sensitive and multidrug-resistant acute lymphoblastic leukemia (ALL) cells. Taken together, this work describes promising colchicine-site-targeting tubulin inhibitors featuring favorable therapeutic effects against ALL and multidrug-resistant cells.
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U2 - 10.1016/j.isci.2019.10.003
DO - 10.1016/j.isci.2019.10.003
M3 - Article
C2 - 31655259
AN - SCOPUS:85073606426
VL - 21
SP - 95
EP - 109
JO - iScience
JF - iScience
SN - 2589-0042
ER -