Electrophilic tuning of the chemoprotective natural product sulforaphane

Young Hoon Ahn, You Sang Hwang, Hua Liu, Xiu Jun Wang, Ying Zhang, Katherine K. Stephenson, Tatiana N. Boronina, Robert N Cole, Albena T. Dinkova-Kostova, Paul Talalay, Philip A. Cole

Research output: Contribution to journalArticle

Abstract

Sulforaphane [1-isothiocyanato-4-(methylsulfinyl)butane], a naturally occurring isothiocyanate derived from cruciferous vegetables, is a highly potent inducer of phase 2 cytoprotective enzymes and can protect against electrophiles including carcinogens, oxidative stress, and inflammation. The mechanism of action of sulforaphane is believed to involve modifications of critical cysteine residues of Keap1, which lead to stabilization of Nrf2 to activate the antioxidant response element of phase 2 enzymes. However, the dithio-carbamate functional group formed by a reversible reaction between isothiocyanate of sulforaphane and sulfhydryl nucleophiles of Keap1 is kinetically labile, and such modification in intact cells has not yet been demonstrated. Here we designed sulforaphane analogs with replacement of the reactive isothiocyanate by the more gentle electrophilic sulfoxythiocarbamate group that also selectively targets cysteine residues in proteins but forms stable thiocarbamate adducts. Twenty-four sulfoxythiocarbamate analogs were synthesized that retain the structural features important for high potency in sulforaphane analogs: the sulfoxide or keto group and its appropriate distance to electrophilic functional group. Evaluation in various cell lines including hepatoma cells, retinal pigment epithelial cells, and keratinocytes as well as in mouse skin shows that these analogs maintain high potency and efficacy for phase 2 enzyme induction as well as the inhibitory effect on lipopolysaccharide-induced nitric oxide formation like sulforaphane. We further show in living cells that a sulfoxythiocarbamate analog can label Keap1 on several key cysteine residues as well as other cellular proteins offering new insights into the mechanism of chemoprotection.

Original languageEnglish (US)
Pages (from-to)9590-9595
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number21
DOIs
StatePublished - May 25 2010

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Biological Products
Cysteine
sulfoxide
Thiocarbamates
Antioxidant Response Elements
Enzyme Induction
Retinal Pigments
Carbamates
Enzymes
Keratinocytes
Vegetables
Carcinogens
Lipopolysaccharides
sulforafan
Hepatocellular Carcinoma
Nitric Oxide
Proteins
Oxidative Stress
Epithelial Cells
Inflammation

ASJC Scopus subject areas

  • General

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Electrophilic tuning of the chemoprotective natural product sulforaphane. / Ahn, Young Hoon; Hwang, You Sang; Liu, Hua; Wang, Xiu Jun; Zhang, Ying; Stephenson, Katherine K.; Boronina, Tatiana N.; Cole, Robert N; Dinkova-Kostova, Albena T.; Talalay, Paul; Cole, Philip A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 21, 25.05.2010, p. 9590-9595.

Research output: Contribution to journalArticle

Ahn, YH, Hwang, YS, Liu, H, Wang, XJ, Zhang, Y, Stephenson, KK, Boronina, TN, Cole, RN, Dinkova-Kostova, AT, Talalay, P & Cole, PA 2010, 'Electrophilic tuning of the chemoprotective natural product sulforaphane', Proceedings of the National Academy of Sciences of the United States of America, vol. 107, no. 21, pp. 9590-9595. https://doi.org/10.1073/pnas.1004104107
Ahn, Young Hoon ; Hwang, You Sang ; Liu, Hua ; Wang, Xiu Jun ; Zhang, Ying ; Stephenson, Katherine K. ; Boronina, Tatiana N. ; Cole, Robert N ; Dinkova-Kostova, Albena T. ; Talalay, Paul ; Cole, Philip A. / Electrophilic tuning of the chemoprotective natural product sulforaphane. In: Proceedings of the National Academy of Sciences of the United States of America. 2010 ; Vol. 107, No. 21. pp. 9590-9595.
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AU - Stephenson, Katherine K.

AU - Boronina, Tatiana N.

AU - Cole, Robert N

AU - Dinkova-Kostova, Albena T.

AU - Talalay, Paul

AU - Cole, Philip A.

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AB - Sulforaphane [1-isothiocyanato-4-(methylsulfinyl)butane], a naturally occurring isothiocyanate derived from cruciferous vegetables, is a highly potent inducer of phase 2 cytoprotective enzymes and can protect against electrophiles including carcinogens, oxidative stress, and inflammation. The mechanism of action of sulforaphane is believed to involve modifications of critical cysteine residues of Keap1, which lead to stabilization of Nrf2 to activate the antioxidant response element of phase 2 enzymes. However, the dithio-carbamate functional group formed by a reversible reaction between isothiocyanate of sulforaphane and sulfhydryl nucleophiles of Keap1 is kinetically labile, and such modification in intact cells has not yet been demonstrated. Here we designed sulforaphane analogs with replacement of the reactive isothiocyanate by the more gentle electrophilic sulfoxythiocarbamate group that also selectively targets cysteine residues in proteins but forms stable thiocarbamate adducts. Twenty-four sulfoxythiocarbamate analogs were synthesized that retain the structural features important for high potency in sulforaphane analogs: the sulfoxide or keto group and its appropriate distance to electrophilic functional group. Evaluation in various cell lines including hepatoma cells, retinal pigment epithelial cells, and keratinocytes as well as in mouse skin shows that these analogs maintain high potency and efficacy for phase 2 enzyme induction as well as the inhibitory effect on lipopolysaccharide-induced nitric oxide formation like sulforaphane. We further show in living cells that a sulfoxythiocarbamate analog can label Keap1 on several key cysteine residues as well as other cellular proteins offering new insights into the mechanism of chemoprotection.

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