Synthetic cyclopenta [b] indoles exhibit antineoplastic activity by targeting microtubule dynamics in acute myeloid leukemia cells

Author(s):Show less -

Vicari, Hugo Passos ^[1] ; Lima, Keli ^[1] ; Gomes, Ralph da Costa ^[2] ; Fernandes, Daniara Cristina ^{[3, 2]} ; Lipreri da Silva, Jean Carlos ^[1] ; Rodrigues Junior, Manoel Trindade ^[2] ; Barroso de Oliveira, Aline Silva ^[2] ; dos Santos, Ricardo Nascimento ^[4] ; Andricopulo, Adriano Defini ^[4] ; Coelho, Fernando ^[2] ; Costa-Lotufo, Leticia Veras ^[1] ; Machado-Neto, Joao Agostinho ^[1]

Affiliation

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Pharmacol, Av Prof Lineu Prestes 1524, BR-05508900 Sao Paulo, SP - Brazil

^[2] Univ Estadual Campinas, Chem Inst, Dept Organ Chem, BR-13083970 Campinas, SP - Brazil

^[3] Inst Fed Educ Ciencia & Tecnol Sao Paulo, BR-15991502 Matao, SP - Brazil

^[4] Univ Sao Paulo, Phys Inst Sao Carlos, BR-13565905 Sao Carlos, SP - Brazil

• Document type: Journal article
• Source: European Journal of Pharmacology; v. 894, MAR 5 2021.
• Web of Science Citations: 0

Abstract

Acute promyelocytic leukemia (APL) is associated with PML-RAR alpha oncogene, which is treated using all-trans retinoic acid (ATRA)-based chemotherapy. However, chemoresistance is observed in 20-30% of treated patients and represents a clinical challenge, raising the importance of the development of new therapeutic options. In the present study, the effects of three synthetic cyclopenta{[}b]indoles on the leukemia phenotype were investigated using NB4 (ATRA-sensitive) and NB4-R2 (ATRA-resistant) cells. Among the tested synthetic cyclopenta{[}b]indoles, compound 2, which contains a heterocyclic nucleus, was the most active, presenting time-dependent cytotoxic activity in the mu M range in APL cells, without cytotoxicity for normal leukocytes, and was selected for further characterization. Compound 2 significantly decreased clonogenicity, increased apoptosis, and caused cell cycle arrest at S and G(2)/M phases in a drug concentration-dependent manner. Morphological analyses indicated aberrant mitosis and diffuse tubulin staining upon compound 2 exposure, which corroborates cell cycle findings. In the molecular scenario, compound 2 reduced STMN1 expression and activity, and induced PARP1 cleavage and H2AX and CHK2 phosphorylation, and modulated CDKN1A, PMAIP1, GADD45A, and XRCC3 expressions, indicating reduction of cell proliferation, apoptosis, and DNA damage. Moreover, in the in vivo tubulin polymerization assay, NB4 and NB4-R2 cells showed a reduction in the levels of polymerized tubulin upon compound 2 exposure, which indicates tubulin as a target of the drug. Molecular docking supports this hypothesis. Taken together, these data indicated that compound 2 exhibits antileukemic effects through disrupting the microtubule dynamics, identifying a possible novel potential antineoplastic agent for the treatment of ATRA-resistant APL. (AU)

• FAPESP's process: 19/01700-2 - Functional investigation of Stathmin 1 in proliferation, differentiation and apoptosis in acute promyelocytic leukemia model
• Grantee: Hugo Passos Vicari
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery
• Grantee: Glaucius Oliva
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 19/23864-7 - Comprehensive analysis of genomic data for identification and validation of novel therapeutic targets involved in cellular cytoskeleton regulation in acute Leukemia
• Grantee: João Agostinho Machado Neto
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/17177-6 - Integrative approach on the sustainable prospection of marine natural products: from diversity to anticancer compounds
• Grantee: Leticia Veras Costa Lotufo
• Support Opportunities: BIOTA-FAPESP Program - Thematic Grants
• FAPESP's process: 17/24993-0 - Investigation of Stathmin 1 and microtubule instability in phenotype of hematological neoplasms
• Grantee: João Agostinho Machado Neto
• Support Opportunities: Regular Research Grants