Zinc-responsive fluorophores
Inventors
Garwin, Seth A. • Que, Emily L. • O'Halloran, Thomas V. • Woodruff, Teresa K.
Assignees
Publication Number
US-12259325-B2
Publication Date
2025-03-25
Expiration Date
2040-09-25
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Abstract
The invention relates generally to compositions and methods for the detection of zinc. Provided herein is a class of zinc-responsive probes with tunable photophysical properties that can be modified for coupling to a solid support or other chemical moieties. In particular, modifications to the 5-position of the BODIPY core allows for alteration of probe properties and functionalities.
Core Innovation
The invention provides a class of zinc-responsive probes based on a BODIPY core with tunable photophysical properties. Modifications at the 5-position of the BODIPY core allow for alteration of probe properties and functionalities, enabling their use in zinc detection and interaction with other chemical moieties or solid supports.
These probes, referred to as ZincBY zinc sensors, can be tailored through their pendant chain at the 5-position, where various substituents such as alkyl, azide, amine, or acetamide are used to introduce different functionalities. This enables the generation of probes with altered zinc affinities, emission properties, and the capability to couple to surfaces or other functional groups.
The problem addressed by the invention is the limitation of existing zinc probes, which often require hydrolysis of toxic protecting groups or application at high concentrations that can negatively impact cell function. The improved BODIPY-based zinc probes described herein can be used at much lower concentrations and offer enhanced specificity, photophysical tunability, ability to conjugate to supports, and compatibility with imaging techniques for monitoring zinc fluctuations and localization.
Claims Coverage
There is one independent claim in this patent, which introduces a method involving a zinc-responsive probe for zinc detection.
Method for detecting zinc using a zinc-responsive probe with a modified BODIPY core
The method comprises contacting a sample with a zinc-responsive probe that includes a BODIPY core modified at the 5-position, where the pendant group R is selected from a wide range of chemical entities such as H, alkyl, substituted alkyl, branched alkyl, substituted branched alkyl, hydroxy, alkoxy, amine, substituted amine, alkylamine, substituted alkylamine, thioalkyl, alkylthioalkyl, azide, cyanide, thioalkyl, ether, ester, thiol, thioether, amino hydroxyl, halogen, ketone, carboxyl, amide, substituted amide, alkylamide, substituted alkylamide, cyano, sulfonyl, carboxy, dialkylphosphine oxide, and combinations thereof. R is not —O—CH3 in some embodiments.
The main inventive feature focuses on a method utilizing a highly customizable zinc-responsive probe with specific structural modifications to the BODIPY core, allowing for tailored zinc detection in various samples.
Stated Advantages
The zinc-responsive probes can be used at much lower concentrations (e.g., nanomolar) compared to existing probes that require micromolar amounts.
Modifications at the 5-position of the BODIPY core allow alteration and optimization of probe properties, including zinc affinity, emission spectrum, and Stokes shift.
Some probes exhibit a large Stokes shift (up to 100 nm), enabling simultaneous imaging with other fluorescent or luminescent probes using a single excitation wavelength.
These probes can be conjugated to a solid support or coupled with other functional moieties, allowing for the creation of extracellular or surface-bound zinc probes.
The probes are zinc-specific and do not require toxic protecting group hydrolysis, minimizing negative impacts on cells.
Probes demonstrate high selectivity for zinc over other biologically relevant metal ions and have stable or minimal pH dependence in the physiological range.
Documented Applications
Detection and imaging of zinc concentration and localization in samples including cells, tissues, organs, or whole animals.
Live-cell imaging to probe static and dynamic zinc pools or zinc fluxes within live cells.
Simultaneous imaging of zinc and other targets (e.g., GFP, tubulin) in living cells or organisms using a single excitation wavelength and minimal spectral overlap.
Conjugation of zinc-responsive probes to solid supports (e.g., glass slides, beads, particles) or chemical moieties (e.g., nucleic acid, peptides, proteins, antibodies, lipids, affinity molecules) for extracellular zinc detection.
Monitoring of zinc during cell cycle progression, fertilization, and other biological processes in mammalian eggs and model organisms (e.g., C. elegans).
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