A plant
dictamine analog, 1-[4-(furo[2,3-b]quinolin-4-ylamino)phenyl]ethanone (CIL-102) has been shown to exert potent anti-
tumor activity. In this study, we examined the mode of interaction of
CIL-102 with
tubulin and unraveled the cellular mechanism responsible for its anti-
tumor activity.
CIL-102 bound to
tubulin at a single site with a dissociation constant ~0.4 μM. Isothermal titration calorimetry revealed that CIL-102-tubulin interaction is highly enthalpy driven and that the binding affords a large negative heat capacity change (ΔC(p) = -790 cal mol(-1) K(-1)) with an enthalpy-entropy compensation. An analysis of the modified Dixon plot suggested that
CIL-102 competitively inhibited the binding of
podophyllotoxin, a
colchicine-binding site agent, to
tubulin. Computational modeling indicated that
CIL-102 binds exclusively at the β-subunit of
tubulin and that
CIL-102 and
colchicine partially share their binding sites on
tubulin. It bound to
tubulin reversibly and the binding was estimated to be ~1000 times faster than that of
colchicine.
CIL-102 potently inhibited the proliferation of MCF-7 cells, induced monopolar spindle formation and multi-nucleation. At half-maximal inhibitory concentration, the spindle microtubules were visibly depolymerized and disorganized.
CIL-102 reduced the inter-polar distances of bipolar mitotic cells indicating that it impaired microtubule-kinetochore attachments. CIL-102-treatment induced apoptosis in MCF-7 cells in association with increased nuclear accumulation of p53 and p21 suggesting that apoptosis is triggered through a p53-p21 dependent pathway. The results indicated that
CIL-102 exerted anti-proliferative activity by disrupting microtubule functions through
tubulin binding and provided important insights into the differential mode of
tubulin binding by
CIL-102 and
colchicine.