Abstract
Melanoma and nonmelanoma skin cancers are among the most prevalent and most lethal forms of skin cancers. To identify new lead compounds with potential anticancer properties for further optimization, in vitro assays combined with in-silico target fishing and docking have been used to identify and further map out the antiproliferative and potential mode of action of molecules from a small library of compounds previously prepared in our laboratory. From screening these compounds in vitro against A375, SK-MEL-28, A431, and SCC-12 skin cancer cell lines, 35 displayed antiproliferative activities at the micromolar level, with the majority being primarily potent against the A431 and SCC-12 squamous carcinoma cell lines. The most active compounds 11 (A431: IC50 = 5.0 μM, SCC-12: IC50 = 2.9 μM, SKMEL-28: IC50 = 4.9 μM, A375: IC50 = 6.7 μM) and 13 (A431: IC50 = 5.0 μM, SCC-12: IC50 = 3.3 μM, SKMEL-28: IC50 = 13.8 μM, A375: IC50 = 17.1 μM), significantly and dose-dependently induced apoptosis of SCC-12 and SK-MEL-28 cells, as evidenced by the suppression of Bcl-2 and upregulation of Bax, cleaved caspase-3, caspase-9, and PARP protein expression levels. Both agents significantly reduced scratch wound healing, colony formation, and expression levels of deregulated cancer molecular targets including RSK/Akt/ERK1/2 and S6K1. In silico target prediction and docking studies using the SwissTargetPrediction web-based tool suggested that CDK8, CLK4, nuclear receptor ROR, tyrosine protein-kinase Fyn/LCK, ROCK1/2, and PARP, all of which are dysregulated in skin cancers, might be prospective targets for the two most active compounds. Further validation of these targets by western blot analyses, revealed that ROCK/Fyn and its associated Hedgehog (Hh) pathways were downregulated or modulated by the two lead compounds. In aggregate, these results provide a strong framework for further validation of the observed activities and the development of a more comprehensive structure–activity relationship through the preparation and biological evaluation of analogs.
Original language | English (US) |
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Article number | e14418 |
Journal | Chemical Biology and Drug Design |
Volume | 103 |
Issue number | 1 |
DOIs | |
State | Published - Jan 2024 |
Funding
The authors acknowledge the financial support of NSF CHE‐1954734 and from the Edward G. Schlieder Educational Foundation (awarded to JF). The following grants and funding partially supported this work: a Start‐up fund from the University of Louisiana at Monroe (ULM) College of Pharmacy (COP), and a Faculty Research Seed grant #5CALHN‐260615 awards from the ULMCOP; a Pilot and a full research project awards from a Louisiana Biomedical Research Network (LBRN)‐IDeA Networks of Biomedical Research Excellence (INBRE) award from the National Institute of General Medical Sciences of the National Institutes of Health (NIGMS/NIH) grant number P2O GM103424‐18, an LBRN‐INBRE‐COBRE Administrative Supplement Award from NIGMS/NIH grant 3P20GM103424‐18S1, and a Louisiana Board of Regents Support Fund grant LEQSF (2021‐24)‐RD‐A‐22 (awarded to JCC). The content is solely the authors' responsibility and does not necessarily represent the official views of the funding agencies. The authors acknowledge the financial support of NSF CHE-1954734 and from the Edward G. Schlieder Educational Foundation (awarded to JF). The following grants and funding partially supported this work: a Start-up fund from the University of Louisiana at Monroe (ULM) College of Pharmacy (COP), and a Faculty Research Seed grant #5CALHN-260615 awards from the ULMCOP; a Pilot and a full research project awards from a Louisiana Biomedical Research Network (LBRN)-IDeA Networks of Biomedical Research Excellence (INBRE) award from the National Institute of General Medical Sciences of the National Institutes of Health (NIGMS/NIH) grant number P2O GM103424-18, an LBRN-INBRE-COBRE Administrative Supplement Award from NIGMS/NIH grant 3P20GM103424-18S1, and a Louisiana Board of Regents Support Fund grant LEQSF (2021-24)-RD-A-22 (awarded to JCC). The content is solely the authors' responsibility and does not necessarily represent the official views of the funding agencies.
Keywords
- ROCK/Fyn and Hedgehog (Hh) pathway as potential targets
- anticancer activity
- apoptosis
- in silico target(s) prediction
- melanoma and nonmelanoma skin cancer cells
ASJC Scopus subject areas
- Biochemistry
- Molecular Medicine
- Pharmacology
- Drug Discovery
- Organic Chemistry