Naphtalene triplet excited state as a probe for the assessment of drug distribution in binary protein systems

[EN] Three drugs containing the naphthalene (NP) chromophore, namely, naproxen (NPX), propranolol (PPN), and cinacalcet (CIN), but with different affinities toward serum albumins (SAs) and alpha-1-acid glycoproteins (AAGs) have been employed for the assessment of drug distribution in binary SA/AAG systems. These three drugs represent an appropriate choice for checking whether a methodology based on transient absorption spectroscopy of a given reporter can be employed for discrimination between different distribution patterns in multicompartmental biological media. Thus, upon laser flash photolysis (LFP) of NPX, PPN, and CIN in the presence or absence of proteins, the NP triplet excited state ((NP)-N-3*) at similar to 420 nm was always detected, although the kinetics of the decay traces was structure- and medium-dependent. In aerated PBS, only a very short triplet lifetime (tau(T)) was found (1-2 mu s). By contrast, in the presence of SAs, two longer triplet lifetimes (5-76 mu s) were observed, ascribed to (NP)-N-3* within site land site II. Upon binding to AAGs, only a long tau(T) (15-47 mu s) was found. When the two proteins were present simultaneously in the same media, fitting of the decay traces was clearly consistent with a distribution of the drug between the different biological compartments and the bulk solution, which correlates well with the known protein affinities of every drug. Experiments were performed in both human (HSA/HAAG) and bovine protein media (BSA/BAAG). The results showed that SAs are the major carriers for NPX; by contrast, PPN binds preferentially to AAGs. An intermediate situation was found for CIN, which presents comparable affinity for both proteins. The results obtained for the two enantiomers of each drug were very similar, although a small stereodifferentiation was observed between the triplet lifetimes in the protein binding sites.
Financial support from the MICINN (Grants CTQ2007-67010 and CTQ2010-14882), from the Generalitat Valenciana (Prometeo Program, Ref 2008/090), from Fundacion Cajamurcia, and from Carlos III Institute of Health (Grant RIRAAF, RETICS program) is gratefully acknowledged.