Tested compounds, previously synthesized, are derivatives of chloroquine, drug commonly
used in the treatment and prevention of malaria. Human serum albumin (HSA) has the role in
transport of endogenous (fatty acids, hormones, bile acids, amino acids) and exogenous
compounds (drug molecules and nutrients). Interaction between tested compounds and HSA
has been studied by fluorescence spectroscopy in phosphate buffered saline (1× PBS, pH 7.4)
[1]. Results show that among tested compounds, all positively charged at pH 7.4, derivatives
with thiophene substructure bind to HSA. Molecular docking studies were used to determine
HSA–compound binding mode.
Fluorescence quenching data were processed using Stern-Volmer (S-V) equation [2]. Almost
linear S-V plot for binding of 1 to HSA (Fig. 1a) indicates single type of quenching mechanism.
Results show that Ksv decreases (20C: (2.60±0.07)×105 M
-1
; 25C: (2.33±0.07)×105 M
-1 and
37C: (2.18±0.08)×105 M
-1
) as temperature increases indicating static quenching mechanism.
Downward curvature in S-V plots of 2 and 3 (Fig. 1b and 1c) indicates that tryptophan residues
are not fully accessible to the drug and that dynamic quenching dominates over static. Fraction
of tryptophan residues that are buried and inaccessible to the quencher and effective
quenching constants can be determined by modified S-V equation. The effective quenching
constant for 2 and 3 increases as temperature increases, this is another indication that dynamic
quenching process is dominant in binding of 2 and 3 to HSA. Effective quenching constants of
all three compounds are in the order of 10 5 M-1, meaning that these compounds can be
effectively carried and stored by HSA in the human body
