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Updated: Feb 25, 2026
Functionalized Spirocyclic Heterocycle Synthesis and Cytotoxicity Assay
Published on: February 9, 2021
Synthesis and evaluation of modified chalcone based p53 stabilizing agents
Sunniya Iftikhar1, Sardraz Khan1, Aishah Bilal2
1Department of Chemistry, SBASSE, Lahore University of Management Sciences, Lahore 54792, Pakistan.
Abstract:
Tumor suppressor protein p53 induces cell cycle arrest and apoptotic cell death in response to various cellular stresses thereby preventing cancer development. Activation and stabilization of p53 through small organic molecules is, therefore, an attractive approach for the treatment of cancers retaining wild-type p53. In this context, a series of nineteen chalcones with various substitution patterns of functional groups including chloro, fluoro, methoxy, nitro, benzyloxy, 4-methyl benzyloxy was prepared using Claisen-Schmidt condensation. The compounds were characterized using NMR, HRMS, IR and melting points. Evaluation of synthesized compounds against human colorectal (HCT116) and breast (CAL-51) cancer cell lines revealed potent antiproliferative activities. Nine compounds displayed GI50 values in the low micromolar to submicromolar range; for example (E)-1-phenyl-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one (SSE14108) showed GI50 of 0.473±0.043µM against HCT116 cells. Further analysis of these compounds revealed that (E)-3-(4-chlorophenyl)-1-phenylprop-2-en-1-one (SSE14105) and (E)-3-(4-methoxyphenyl)-1-phenylprop-2-en-1-one (SSE14106) caused rapid (4 and 8-h post-treatment) accumulation of p53 in HCT116 cells similar to its induction by positive control, Nutlin-3. Such activities were absent in 3-(4-methoxyphenyl)propiophenone (SSE14106H2) demonstrating the importance of conjugated ketone for antiproliferative and p53 stabilizing activity of the chalcones. We further evaluated p53 levels in the presence of cycloheximide (CHX) and the results showed that the p53 stabilization was regulated at post-translational level through blockage of its degradation. These chalcones can, therefore, act as fragment leads for further structure optimization to obtain more potent p53 stabilizing agents with enhanced anti-proliferative activities.
Insights
New chalcone compounds were synthesized and tested for their ability to activate the tumor suppressor protein p53. Several compounds showed potent anticancer activity by stabilizing p53, offering potential leads for cancer drug development.
Area of Science:
- Medicinal Chemistry
- Organic Synthesis
- Cancer Biology
Background:
- The tumor suppressor protein p53 is crucial in preventing cancer by inducing cell cycle arrest and apoptosis.
- Activating wild-type p53 using small molecules is a promising strategy for cancer treatment.
Purpose of the Study:
- To synthesize and evaluate a series of novel chalcone derivatives for their antiproliferative activity and ability to stabilize p53.
- To explore the structure-activity relationship of these chalcones in cancer cells.
Main Methods:
- Nineteen chalcone derivatives were synthesized via Claisen-Schmidt condensation and characterized using spectroscopic methods (NMR, HRMS, IR) and melting point analysis.
- Antiproliferative activity was assessed against human colorectal (HCT116) and breast (CAL-51) cancer cell lines, determining GI50 values.
- p53 stabilization was evaluated by Western blot analysis in the presence and absence of cycloheximide (CHX).
Main Results:
- Nine chalcone compounds exhibited potent antiproliferative activity, with GI50 values in the low micromolar to submicromolar range.
- Specific compounds, (E)-3-(4-chlorophenyl)-1-phenylprop-2-en-1-one and (E)-3-(4-methoxyphenyl)-1-phenylprop-2-en-1-one, rapidly accumulated p53 in HCT116 cells.
- p53 stabilization was confirmed to be regulated at the post-translational level by inhibiting protein degradation, and the conjugated ketone moiety was essential for activity.
Conclusions:
- The synthesized chalcones demonstrate significant potential as anticancer agents by stabilizing the p53 protein.
- These compounds serve as valuable fragment leads for developing more potent p53-stabilizing drugs with enhanced anti-proliferative effects.
