Invention:
A protected small molecule probe is delivered intracellularly and subsequently releases an aryl diazonium ion for use as a bioconjugation agent or drug delivery agent. This technology enables a novel chemical probe for observing intact biological cells. It builds on previous IP developed by Jewett laboratory for protection, as a class of chemistry enabling stable compounds that then could release reactive species at biological pH, reducing intracellular environment, or via enzymatic release to elucidate biochemistry. This invention comprises a class of chemistry and method of use for small molecules with functional groups such that four conditions are met:
- To protect the diazonium ion during delivery
- Provide for cellular uptake
- Provide for intracellular release of the diazonium ion
- Optionally provide for the protein binding within the cell
Background:
Selectivity in biological systems comes from a complex interplay of location, interactions, and reactivity. Covalent small-molecule probes offer great potential in the development of chemical tools to study intracellular proteins. A challenge within this area is in garnering selectivity associated with location. For example, in order to gain accessibility to intracellular proteins, small-molecule labeling strategies have relied on working with cell lysate, yet key contextual interactions associated with localization within the cell are lost during lysis.
Aryl diazonium ions are known for their selective reactivity with the electron-rich aromatic tyrosine side chain. But aryl diazonium ions for use as probes have suffered from a lack of deliverability since they have short half-lives and are generally unstable.
The present invention provides probes, e.g., protected triazabutadiene probes, that selectively release benzene diazonium ions (BDIs) intracellularly, providing a tool for cellular studies in intact biological systems, e.g., a means for accessing and/or labeling intracellular proteins or molecules prior to cell lysis.
Applications:
- Protein labeling
- Bioconjugation
- Drug discovery
- Life science research
- Tyrosine studies
- Histidine studies
Advantages:
- Enables in cellulo studies
- Covalent binding
- Tunable chemistry for selectivity and delivery
