Open Access
ARTICLE
Discovery of Two Novel Pyrazole Derivatives as Anticancer Agents Targeting Tubulin Polymerization and MAPK Signaling Pathways
Denisse A. Gutierrez*, Elisa Robles-Escajeda, Jose A. Lopez-Saenz, Robert A. Kirken, Edgar A. Borrego, Ana P. Betancourt, Soumya Nair, Sourav Roy, Armando Varela-Ramirez, Renato J. Aguilera*
Border Biomedical Research Center and Department of Biological Sciences, The University of Texas at El Paso, El Paso, TX, USA
* Corresponding Author: Denisse A. Gutierrez. Email: 
; Renato J. Aguilera. Email: 
(This article belongs to the Special Issue: Discovery of a Potent Antitumor Agent: Mechanistic Insights and Therapeutic Potential)
Oncology Research https://doi.org/10.32604/or.2026.074945
Received 22 October 2025; Accepted 30 January 2026; Published online 21 February 2026
Abstract
Objectives: Drug resistance is the major determinant of chemotherapy failure, leading to relapse and tumor progression, demonstrating the urgent need for novel antineoplastic drugs. This study aimed to evaluate the anticancer potential of two novel pyrazole derivatives, P3C.1 and P3C.2, and to elucidate their mechanism of action in cancer cells. Methods: The cytotoxicity of the compounds was evaluated across 27 different cancer cell lines via a nuclear staining assay. Subsequent flow cytometric and biochemical analyses were performed to assess reactive oxygen species (ROS) generation, apoptosis induction, mitochondrial integrity, and cell cycle progression. Additional studies included transcriptome analyses and immunoassays to characterize the molecular mechanisms underlying drug activity. Results: Two novel pyrazole derivatives, P3C.1 and P3C.2, were identified with potent cytotoxicity on a variety of cancer cell lines. Among the adherent cell lines tested, the triple-negative breast cancer (TNBC) cell line MDA-MB-231 exhibited the highest sensitivity to both compounds and was therefore selected for further experimentation. In vitro assays demonstrated that both compounds induced ROS generation, mitochondrial membrane depolarization, cell cycle arrest and apoptosis. Whole-transcriptome sequencing of P3C.1 and P3C.2-treated MDA-MB-231 and two lymphoblastic leukemia cell lines revealed four genes in common associated with cell signaling and membrane dynamics. Connectivity Map (CMAP) database comparisons of shared genes for each cancer subtype revealed a strong similarity between the two compounds with tubulin inhibitors, and subsequent assays confirmed that these compounds act as microtubule-disrupting agents. Moreover, protein phosphorylation analysis indicated that both compounds induced hyperphosphorylation of JNK, and ERK1/2, along with hypophosphorylation of p38 kinases. Conclusions: P3C.1 and P3C.2 emerged as promising anti-breast cancer agents with dual mechanisms of action involving microtubule disruption and altered kinase signaling, leading to induction of apoptosis.
Keywords
Pyrazoles; cytotoxicity; triple-negative breast cancer (TNBC); apoptosis; tubulin polymerization inhibition; phosphorylation
Login
Register
Submit a Paper
Propose a Special lssue
Download PDF
Supplementary Material File
Downloads
Citation Tools
