Fluorinated o-aminophenol derivatives for measurement of intracellular pH

The simple 2-aminophenol group which serves as a building block for many cationic indicators has been modified to yield a series of pH sensitive probes. This approach is based on the replacement of one of the N-acetate groups of the chelator APTRA (o-aminophenol N,N,O-triacetate) by an N-ethyl group. The resulting series of (N-ethylamino)phenol (NEAP) compounds exhibit pK values in the physiological range and negligible affinity for physiological levels of other ions. Three fluorinated analogs have been prepared: N-ethyl-5-fluoro-2-aminophenol N,O-diacetate (5F NEAP), N-ethyl-2-((2-fluoro-4-carboxybenzyl)oxy)-4-fluoroaniline-N- acetic acid (5F NEAP-2), and 1-(2-(N-ethylamino)-5-fluorophenoxy)-2-(2-fluoro-4- aminophenoxy)ethane-N,N',N'-triacetic acid (5F NEAP-3). These derivatives exhibit total titration shifts of approximately 11 ppm. NEAP-2 and NEAP-3 contain an additional fluorine to serve as an internal chemical shift reference, and NEAP-3, the most highly charged analog prepared, was designed in order to minimize leakage.