Fluorogenic H2O2 Probe Using Mislow-Evans Rearrangement

Solution Overview

Problem

Existing chemical probes for hydrogen peroxide (H2O2) in biological systems are slow to produce fluorescence signals, lacking the specificity and temporal resolution needed for precise detection, particularly due to reliance on boronate ester functionality that requires the presence of the hydroperoxide anion (HOO−), which is scarce under biological conditions.

Innovation Solution

A compound utilizing a seleno-Mislow-Evans rearrangement integrated with a spontaneous acetal hydrolysis mechanism, allowing rapid detection of H2O2 through a fluorogenic switch, which is more reactive with H2O2 than other reactive oxygen and nitrogen species (ROS and RNS).

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If boronate ester functionality is used for H2O2 detection, then selectivity for H2O2 is improved, but reaction kinetics deteriorate (slow fluorescence signal production)

Engineering Contradiction:

ImproveselectivityVSAvoidreaction kinetics

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent changes the chemical functionality from boronate ester to selenite ester, altering the reaction mechanism from requiring hydroperoxide anion to reacting with neutral H2O2. This parameter change in chemical composition and reactivity resolves the contradiction by achieving both selectivity and fast kinetics simultaneously.

Inventive Principle:

Principle #35Parameter changes

2Measurement precision

If boronate ester probes are used, then H2O2 detection is achieved, but temporal resolution deteriorates (takes ~30 min to produce fluorescence signals)

Engineering Contradiction:

Improvedetection capabilityVSAvoidsignal production time

Core Design Contradiction:

Measurement precisionVSLoss of time

Solution Approach 1:

The patent changes the chemical functionality from boronate ester to selenite ester, altering the reaction mechanism from requiring hydroperoxide anion to reacting with neutral H2O2. This parameter change in chemical composition and reactivity resolves the contradiction by achieving both selectivity and fast kinetics simultaneously.

Inventive Principle:

Principle #35Parameter changes

3Measurement precision

If probes requiring hydroperoxide anion are used, then selectivity is maintained, but detection speed deteriorates (abundance of HOO− is very low ≈0.1% of H2O2)

Engineering Contradiction:

ImproveselectivityVSAvoiddetection speed

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent changes the chemical functionality from boronate ester to selenite ester, altering the reaction mechanism from requiring hydroperoxide anion to reacting with neutral H2O2. This parameter change in chemical composition and reactivity resolves the contradiction by achieving both selectivity and fast kinetics simultaneously.

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The compound enables rapid and selective detection of H2O2 with improved kinetics, capable of visualizing and quantifying H2O2 presence in cells and biological samples, including zebrafish wound-healing models, with high sensitivity and specificity.

Implementation Method 1

A compound utilizing a seleno-Mislow-Evans rearrangement integrated with a spontaneous acetal hydrolysis mechanism, allowing rapid detection of H2O2

Methodology Applied

Scientific EffectMislow-Evans rearrangement:

Implementation Method 2

A compound utilizing a seleno-Mislow-Evans rearrangement integrated with a spontaneous acetal hydrolysis mechanism, allowing rapid detection of H2O2 through a fluorogenic switch

Methodology Applied

Scientific EffectAcetal hydrolysis: Hydrolysis

Implementation Method 3

illuminating the cell, tissue, organ, or organism with light including, or at an excitation wavelength for, the reaction product of the compound with H2O2, and detecting fluorescent emission from the reaction product

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Data Source

PatentUS12540150B1Fluorogenic probe using a Mislow-Evans rearrangement for real-time imaging of hydrogen peroxide

Publication Date: 2026.02.03 UNIV OF PITTSBURGH OF THE COMMONWEALTH SYST OF HIGHER EDUCATION

AI summary

Provided herein is a fluorogenic probe that selectively undergoes a [2,3]-sigmatropic rearrangement (seleno-Mislow-Evans rearrangement) with H2O2, followed by an acetal hydrolysis, to produce a green fluorescent molecule in seconds. Also provided herein is a method of identifying or visualizing the presence of H2O2 in a cell, tissue, organ, or organism that includes contacting the cell, tissue, organ, or organism with the fluorogenic probe. Also provided herein is a method of identifying or quantifying the presence of H2O2 in a sample that includes adding to or mixing in the sample the fluorogenic probe.