Aminomethylphosphonate and 2-aminoethylphosphonate as31P-NMR pH markers for extracellular and cytosolic spaces in the isolated perfused rat liver

Vidal, Giovanni; Thiaudière, Eric; Canioni, Paul; Gallis, Jean‐Louis

doi:10.1002/1099-1492(200008)13:5<289::aid-nbm647>3.0.co;2-g

Cited by 10 publications

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References 27 publications

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“…To illustrate, the methylphosphonate p K a = 7.36, with Δ δ ab = 3.88 ppm at 20 °C in a Krebs–Henseleit (KH) buffer is currently used in organ perfusion experiments [ 19 ]. Further targets were α-, β- and γ-substituted linear aminophosphonic acids ( Figure 1 B) whose Δ δ ab values, however, did not exceed 3.4 ppm [ 22 ], but showed relatively weak toxicity and had lower p K a s suitable for probing the acidic domains in cells, tumors, and perfused organs [ 23 , 24 , 25 , 26 ]. Since in all these above compounds, protonation always occurs at the phosphorus first, it was then speculated that scaffolds bearing a non-ionizable P -atom (e.g., a phosphonate) and a protonable α-amino group (typically, a secondary amine) would offer considerable advantages in terms of Δ δ ab and p K a modulation due to different inductive and steric effects.…”

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confidence: 99%

Novel Sterically Crowded and Conformationally Constrained α-Aminophosphonates with a Near-Neutral pKa as Highly Accurate 31P NMR pH Probes. Application to Subtle pH Gradients Determination in Dictyostelium discoideum Cells

et al. 2022

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In order to discover new 31P NMR markers for probing subtle pH changes (<0.2 pH unit) in biological environments, fifteen new conformationally constrained or sterically hindered α-aminophosphonates derived from diethyl(2-methylpyrrolidin-2-yl)phosphonate were synthesized and tested for their pH reporting and cytotoxic properties in vitro. All compounds showed near-neutral pKas (ranging 6.28–6.97), chemical shifts not overlapping those of phosphorus metabolites, and spectroscopic sensitivities (i.e., chemical shifts variation Δδab between the acidic and basic forms) ranging from 9.2–10.7 ppm, being fourfold larger than conventional endogenous markers such as inorganic phosphate. X-ray crystallographic studies combined with predictive empirical relationships and ab initio calculations addressed the inductive and stereochemical effects of substituents linked to the protonated amine function. Satisfactory correlations were established between pKas and both the 2D structure and pyramidalization at phosphorus, showing that steric crowding around the phosphorus is crucial for modulating Δδab. Finally, the hit 31P NMR pH probe 1b bearing an unsubstituted 1,3,2-dioxaphosphorinane ring, which is moderately lipophilic, nontoxic on A549 and NHLF cells, and showing pKa = 6.45 with Δδab = 10.64 ppm, allowed the first clear-cut evidence of trans-sarcolemmal pH gradients in normoxic Dictyostelium discoideum cells with an accuracy of <0.05 pH units.

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