Reactivity toward Thiols and Cytotoxicity of 3-Methylene-2-oxindoles, Cytotoxins from Indole-3-acetic Acids, on Activation by Peroxidases
Abstract
Oxidation of indole-3-acetic acid and its derivatives by peroxidases such as that from horseradish produces many products, including 3-methylene-2-oxindoles. These have long been associated with biological activity, but their reactivity has not been characterized. We have previously demonstrated the potential value of substituted indole acetic acids and horseradish peroxidase as the basis for targeted cancer therapy, since the compounds are of low cytotoxicity until oxidized, when high cytotoxicity is observed; the combination of prodrug and enzyme depletes intracellular thiols. In this study, 3-methylene-2-oxindole and derivatives substituted in the 4-, 5-, or 6-position with methyl, F, or Cl have been synthesized and their reactivity toward representative thiol nucleophiles (glutathione, cysteine, and a cysteinyl peptide) measured using stopped-flow kinetic spectrophotometry. Rate constants were in the range approximately 2 x 10(3) to 2 x 10(4) M(-)(1) s(-)(1) at pH 7.4, 25 degrees C, implying a lifetime of a few tens of milliseconds for these methylene oxindoles in the cellular environment and diffusion distances of a few micrometers. As expected, halogen substitution decreased the rate of production of the methylene oxindoles on treatment of horseradish peroxidase. The cytotoxicities of the compounds were measured using Chinese hamster V79 fibroblast-like cells in vitro. The halogen-substituted derivatives were much more cytotoxic than the 5-methyl analogue or the parent (unsubstituted) compound, consistent with the trends in rate constant for reaction with the thiols. The results show that the cytotoxic response in the prodrug (indole acetic acid) and enzyme (horseradish peroxidase) system reflects the reactivity of methylene oxindoles toward nucleophiles much more than the rate of generation of the oxindoles, and helps explain the possible advantages of 5-fluoroindole-3-acetic acid compared to IAA as a lead compound for investigation in targeted cancer therapy.
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Myeloperoxidase can oxidize 3-IAA, inducing toxic products such as 3-methylene-2-oxindole (MOI).
“MPO can oxidize 3-IAA, inducing toxic products such as 3-methylene-2-oxindole (MOI)24,25.”
From Microbiota-derived 3-IAA influences chemotherapy efficacy in pancreatic cancer (2023) · cites this paper as [25]
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Microbiota-derived 3-IAA influences chemotherapy efficacy in pancreatic cancer
Tintelnot, J., Xu, Y., Lesker, T. R., Schönlein, M., Konczalla, L., Giannou, A. D., Pelczar, P., Kylies, D., Puelles, V. G., Bielecka, A. A., et al. (2023) · Nature · doi:10.1038/s41586-023-05728-y · Cited by 478 · see citing works
- Authors
- Folkes, L. K., Rossiter, S., & Wardman, P.
- Affiliations
- Mount Vernon Hospital (GB)
- Corresponding author
- Wardman, Peter
- Venue
- Chemical Research in Toxicology
- Publisher
- American Chemical Society (ACS)
- Published
- 2002-05-21
- Volume
- 15
- Issue
- 6
- Pages
- 877-882
- ISSN
- 0893-228X (ISSN-L)
- 1520-5010
- OpenAlex
- W2007449478
- Cited by
- 12
- References
- 20
- Citations by year
- 2026: 1 · 2023: 1 · 2021: 1 · 2013: 1
- FWCI
- 1.22
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Chemical Research in Toxicology (publishedVersion)
- PubMed (publishedVersion)
- University of Hertfordshire Research Archive (University of Hertfordshire) (acceptedVersion)
- University of Hertfordshire Research Archive (University of Hertfordshire) (submittedVersion)
- Topics
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Toxicology › Bioactive Compounds and Antitumor Agents
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis and biological activity
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- indole-3-acetic acid, horseradish peroxidase, thiol reactivity, glutathione, cysteine, halogen substitution, prodrugs, targeted cancer therapy, cytotoxicity, reaction kinetics
- MeSH terms
- Methanol, Animals, Biotransformation, Cell Survival, Cells, Cultured, Cricetulus, Cysteine, Cytotoxins, Fibroblasts, Glutathione, Cricetinae, Horseradish Peroxidase, Indoleacetic Acids, Indoles, Kinetics, Lung, Oxidation-Reduction, Peptides, Prodrugs, Sulfhydryl Compounds
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