Plumbagin‐Serum Albumin Interaction: Spectral, Electrochemical, Structure‐Binding Analysis, Antiproliferative and Cell Signaling Aspects with Implications for Anticancer Therapy

Plumbagin (5‐hydroxy‐2‐methyl‐1,4‐naphthoquinone) is a small molecule with potent anticancer activity. Like other 1,4‐naphthoquinones, it exhibits electrophilic reactivity towards biological nucleophiles. We demonstrate that plumbagin and structurally related 1,4‐naphthoquinones with at least one unsubstituted quinoid carbon (C2 or C3) bind to albumin, an ubiquitously present nucleophile, with minimum recovery of free drug. Extraction recovery of plumbagin from albumin in solution showed one‐phase exponential decline with a half‐live of 9.3 min at 10 μmol/L. In the presence of albumin, plumbagin exhibited instant changes in UV/Vis absorption bands. Electrochemical analysis using cyclic voltammetry showed a decrease in redox peak currents over time until electro‐inactivity, thus suggesting the formation of a supramolecular adduct inaccessible for electron transfer. The adduct inhibited cell growth and caused cell‐cycle arrest of prostate cancer cells, in part by decreasing levels of the cell‐cycle regulator RBBP. The conjugate displayed similar cellular effects to those described for plumbagin, such as decreased levels of androgen receptor and protein kinase C epsilon. The effect of plumbagin‐albumin on cancer cells was species‐specific, suggesting a receptor‐mediated mechanism. Furthermore, it was blocked by cathepsin inhibitor pepstatin A, indicating that lysosomal degradation is involved in the processing of plumbagin‐albumin adduct. The spontaneously formed adduct of plumbagin with serum albumin is likely to mediate the biological activities of plumbagin in vivo and to fundamentally influence its pharmacodynamics. Stuck on you: Plumbagin, a clinical stage anticancer drug, spontaneously forms an adduct with serum albumin. The plumbagin‐albumin complex inhibits cancer cell proliferation through cell‐cycle arrest, and similar cellular effects are exerted by free plumbagin. The propensity to bind to serum albumin is likely to profoundly affect the pharmacodynamics of plumbagin.