Identification of a Potent Cytotoxic Pyrazole with Anti-Breast Cancer Activity That Alters Multiple Pathways

Abstract

In this study, we identified a novel pyrazole-based derivative (P3C) that displayed potent cytotoxicity against 27 human cancer cell lines derived from different tissue origins with 50% cytotoxic concentrations (CC50) in the low micromolar and nanomolar range, particularly in two triple-negative breast cancer (TNBC) cell lines (from 0.25 to 0.49 µM). In vitro assays revealed that P3C induces reactive oxygen species (ROS) accumulation leading to mitochondrial depolarization and caspase-3/7 and -8 activation, suggesting the participation of both the intrinsic and extrinsic apoptotic pathways. P3C caused microtubule disruption, phosphatidylserine externalization, PARP cleavage, DNA fragmentation, and cell cycle arrest on TNBC cells. In addition, P3C triggered dephosphorylation of CREB, p38, ERK, STAT3, and Fyn, and hyperphosphorylation of JNK and NF-kB in TNBC cells, indicating the inactivation of both p38MAPK/STAT3 and ERK1/2/CREB signaling pathways. In support of our in vitro assays, transcriptome analyses of two distinct TNBC cell lines (MDA-MB-231 and MDA-MB-468 cells) treated with P3C revealed 28 genes similarly affected by the treatment implicated in apoptosis, oxidative stress, protein kinase modulation, and microtubule stability.

Synthesis of Novel Pyrazole‐Oxindole Conjugates with Cytotoxicity in Human Cancer Cells via Apoptosis

“…Cell suspensions were immediately examined by flow cytometry and 100,000 events were acquired for a well‐defined cell cycle distribution profile. Data was analyzed via the Kaluza 1.3 Flow Cytometry Software [31,32] …”

Identification of a Unique Cytotoxic Thieno[2,3-c]Pyrazole Derivative with Potent and Selective Anticancer Effects In Vitro

“…needed to kill 50% of the cell population at a given time point. From the percentage of cell death for each technical replicate at a concentration above and below 50% cell death, CC 50 for each cell line was calculated via linear interpolation and averaged to obtain the reported values [ 7 , 30 , 31 ].…”

“…In these experiments, 5 × 10 6 HL-60 cell samples were collected in microcentrifuge tubes and treated with a sub-CC 50 dose, 0.01 μM or 0.1 μM, of Tpz-1 and incubated for 3 h at 37 °C. The cells in each sample were lysed, their protein concentrations determined, and Multiplex Luminex assays were performed following the same procedure as the experimental compound P3C [ 7 ]. Acquisition and data analysis were performed using Luminex xPONENT 3.1 software.…”

“…Pyrazole derivatives have been approved to treat various leukemias, lymphomas, and other cancers in the United States and China [ 3 ]. In addition, numerous experimental pyrazoles have shown antiproliferative activity and apoptosis against several cancer types in vitro [ 6 , 7 , 8 ]. In contrast, thiophene is a 5-membered heterocycle with four carbon atoms and one sulfur ( Figure 1 ).…”

A virus changed my life

“…As director of our Cellular Characterization Core and Biorepository facility of the Border Biomedical Research Center, which is funded by Research Centers at Minority Institutions, I was able to build a “state-of-the-art” drug-screening facility to continue searching for anticancer agents. Using this facility, we discovered several novel compounds with potent activity against a variety of cancer cell lines ( Santiago-Vazquez et al , 2014 ; Gutierrez et al , 2019, 2022 ). Our group also determined that a known antimalarial drug has potent anticancer activity and is a topoisomerase II inhibitor ( Villanueva et al ., 2018, 2021 ).…”