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Biomolecules Jan 2023Tubulin is a protein that plays a critical role in maintaining cellular structure and facilitating cell division. Inhibiting tubulin polymerization has been shown to be...
Tubulin is a protein that plays a critical role in maintaining cellular structure and facilitating cell division. Inhibiting tubulin polymerization has been shown to be an effective strategy for inhibiting the proliferation of cancer cells. In the past, identifying compounds that could inhibit tubulin polymerization has required the use of in vitro assays utilizing purified tubulin or immunofluorescence of fixed cells. This study presents a novel approach for identifying tubulin polymerization inhibitors using a CRISPR-edited cell line that expresses fluorescently tagged β-tubulin and a nuclear protein, enabling the visualization of tubulin polymerization dynamics via high-content imaging analysis (HCI). The cells were treated with known tubulin polymerization inhibitors, colchicine, and vincristine, and the resulting phenotypic changes indicative of tubulin polymerization inhibition were confirmed using HCI. Furthermore, a library of 429 kinase inhibitors was screened, resulting in the identification of three compounds (ON-01910, HMN-214, and KX2-391) that inhibit tubulin polymerization. Live cell tracking analysis confirmed that compound treatment leads to rapid tubulin depolymerization. These findings suggest that CRISPR-edited cells with fluorescently tagged endogenous β-tubulin can be utilized to screen large compound libraries containing diverse chemical families for the identification of novel tubulin polymerization inhibitors.
Topics: Humans; Tubulin; Tubulin Modulators; Histones; Polymerization; Clustered Regularly Interspaced Short Palindromic Repeats; Cell Line; Antineoplastic Agents; Cell Proliferation; Cell Line, Tumor; Molecular Structure
PubMed: 36830618
DOI: 10.3390/biom13020249 -
Journal of Medicinal Chemistry Jan 2020Inhibiting/disturbing the RAS/RAF pathway may benefit the treatment of cancer and overcome the resistance. Utilizing such a pathway as the target, nine...
Platinum-Based Modification of Styrylbenzylsulfones as Multifunctional Antitumor Agents: Targeting the RAS/RAF Pathway, Enhancing Antitumor Activity, and Overcoming Multidrug Resistance.
Inhibiting/disturbing the RAS/RAF pathway may benefit the treatment of cancer and overcome the resistance. Utilizing such a pathway as the target, nine styrylbenzylsulfone derivatives generated from the platinum-based modification of the side chain of Rigosertib were designed. Among them, compound showed the most potent antitumor activity in vitro with IC values at the nanomolar level against the tested tumor cell lines and 1000-fold higher than cisplatin against the multidrug resistant cells (A549/CDDP, A549/DOX, and SKOV-3/CDDP cells), while it showed only moderate cytotoxicity against normal cells (HEUVC cells). Compound could clearly disturb signaling transduction between RAS and CRAF by directly bonding to CRAF and inhibit CRAF activation. Besides, the enhanced intracellular platinum level made more potent than cisplatin in DNA damage, reactive oxygen species accumulation, and mitochondrial membrane potential decrease. Moreover, induced apoptosis by the endogenous pathway and efficiently inhibited tumor growth in the A549 xenograft model without side effects.
Topics: Animals; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Proliferation; Coordination Complexes; DNA Damage; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Female; G2 Phase Cell Cycle Checkpoints; Glycine; Human Umbilical Vein Endothelial Cells; Humans; Mice, Inbred BALB C; Neoplasms; Platinum; Proto-Oncogene Proteins c-raf; Reactive Oxygen Species; Signal Transduction; Sulfones; Xenograft Model Antitumor Assays; ras Proteins
PubMed: 31820986
DOI: 10.1021/acs.jmedchem.9b01223 -
Cancer Discovery Jun 2016Rigosertib acts as a RAS mimetic, binding to the RAS-binding domain of multiple RAS effector proteins.
Rigosertib acts as a RAS mimetic, binding to the RAS-binding domain of multiple RAS effector proteins.
Topics: Animals; Antineoplastic Agents; Glycine; Humans; Molecular Mimicry; Mutation; Protein Binding; Protein Interaction Domains and Motifs; Protein Kinase Inhibitors; Signal Transduction; Sulfones; ras Proteins
PubMed: 27150538
DOI: 10.1158/2159-8290.CD-RW2016-085 -
Blood Cancer Journal Dec 2017
Randomized Controlled Trial
Validation of a post-hypomethylating agent failure prognostic model in myelodysplastic syndromes patients treated in a randomized controlled phase III trial of rigosertib vs. best supportive care.
Topics: Antineoplastic Agents; Azacitidine; Decitabine; Drug Resistance, Neoplasm; Glycine; Humans; Kaplan-Meier Estimate; Myelodysplastic Syndromes; Prognosis; Sulfones; Treatment Outcome
PubMed: 29238044
DOI: 10.1038/s41408-017-0018-7 -
Cell Cycle (Georgetown, Tex.) Jun 2017For almost a decade, there has been much interest in the development of chemical inhibitors of Polo-like kinase 1 (Plk1) protein interactions. Plk1 is a master regulator...
For almost a decade, there has been much interest in the development of chemical inhibitors of Polo-like kinase 1 (Plk1) protein interactions. Plk1 is a master regulator of the cell division cycle that controls numerous substrates. It is a promising target for cancer drug development. Inhibitors of the kinase domain of Plk1 had some success in clinical trials. However, they are not perfectly selective. In principle, Plk1 can also be inhibited by interfering with its protein interaction domain, the Polo-Box Domain (PBD). Selective chemical inhibitors of the PBD would constitute tools to probe for PBD-dependent functions of Plk1 and could be advantageous in cancer therapy. The discovery of Poloxin and thymoquinone as PBD inhibitors indicated that small, cell-permeable chemical inhibitors could be identified. Other efforts followed, including ours, reporting additional molecules capable of blocking the PBD. It is now clear that, unfortunately, most of these compounds are non-specific protein alkylators (defined here as groups covalently added via a carbon) that have little or no potential for the development of real Plk1 PBD-specific drugs. This situation should be minded by biologists potentially interested in using these compounds to study Plk1. Further efforts are needed to develop selective, cell-permeable PBD inhibitors.
Topics: Alkylation; Antineoplastic Agents; Benzoates; Benzoquinones; Cell Cycle Proteins; Glycine; Humans; Mitosis; Neoplasms; Protein Kinase Inhibitors; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins; Quinones; Sulfones; Polo-Like Kinase 1
PubMed: 28521657
DOI: 10.1080/15384101.2017.1325043