Patent · US Active

Thiol-based fluorescent probe for reactive species

US11339175B2 · kind B2 · utility

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18Claims
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Key dates

Filing dateNov 6, 2019
Grant dateMay 24, 2022
Priority date
Expiry dateFeb 3, 2040

Classification

  • Technology area (CPC Y)Emerging Cross-Sectional Technologies
  • CPC primaryY10T436/177692
  • WIPO fieldMeasurement
  • WIPO sectorInstruments

Abstract

Detection of nitroxyl (HNO), the transient one-electron reduced form of nitric oxide, is a significant challenge owing to its high reactivity with biological thiols (rate constants as high as 109M−1 s−1). Reported herein is a new thiol-based HNO-responsive trigger that can compete against reactive thiols for HNO. This process forms an N-hydroxysulfenamide intermediate which cyclizes to release a masked fluorophore leading to fluorescence enhancement. To ensure a rapid cyclization step, the disclosed design capitalizes on two established physical organic phenomena: the alpha-effect and the Thorpe-Ingold effect. Using this new trigger, NitroxylFluor was developed; a selective HNO-responsive fluorescent probe. Treatment of NitroxylFluor with an HNO donor results in a 16-fold turn-on. This probe also exhibits excellent selectivity over various reactive nitrogen, oxygen, and sulfur species and efficacy in the presence of thiols (e.g., glutathione in mM concentrations). Also, live cell imaging of HNO using NitroxylFluor was performed.

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