Assessment of the complexation degree of camptothecin derivatives and cyclodextrins using spectroscopic and separative methodologies

Abstract

The complexation of camptothecin and homocamptothecin derivatives, topoisomerase I inhibitors, with two cyclodextrins (CDs) of pharmaceutical interest (native and hydroxypropylated β-CD) was studied at pH 3.5 and 6. In a first step, the affinity order of the six compounds studied for the β-CD and HP-β-CD was evaluated in HPLC using immobilized stationary phases [Cyclobond I 2000 (β-CD) and Cyclobond I 2000 RSP (HP-β-CD)]. In a second step, the apparent binding constants of the 12 complexes studied were determined at both pH by HPLC using Scott’s method with CD as a chiral additive. The 1:1 stoichiometry of the complex formed between HP-β-CD and the homocamptothecin derivative elomotecan ( R)-6 was established by fluorescence spectroscopy using the continuous variation method developed by Job and ESI-MS. Complementary investigations were achieved for topotecan ( S)-3 and elomotecan ( R)-6 using CE. Further studies provided similar conclusions concerning affinity of all the derivatives studied for both CDs: that is, a slightly larger affinity was observed for HP-β-CD with respect to β-CD, except for ( S)-3. For ( S)-3, this affinity increase with pH, in the range studied.