Ciliobrevin D is a AAA+ ATPase motor cytoplasmic dynein inhibitor. Ciliobrevin D inhibits Hedgehog (Hh) signaling and primary cilia formation and it also inhibits dynein-dependent microtubule gliding and ATPase activity in vitro.

Ciliobrevin D reversibly inhibits melanosome aggregation, however, the non-cilia-disrupting derivative had no discernible effect at comparable doses. Cells treated with Ciliobrevin D exhibits abnormal (unfocused, multipolar, or collapsed) spindles with disrupted γ-tubulin localization in NIH-3T3 cells. Ciliobrevin D addition also reversibly disrupts the pre-formed spindles of metaphase-arrested cells and reduces overall microtubule levels and it similarly abrogates the movement of peroxisomes in Drosophila S2 cells[1].

In the testis in vivo, Knockdown of Dync1h1 or inactivation of dynein 1 by Ciliobrevin D perturbs spermatogenesis. The use of Ciliobrevin D to inactivate dynein 1 in the testis in vivo perturbs MT organization through changes in the spatial expression of EB1, perturbs F-actin organization, and perturbs distribution of adhesion protein complexes at the BTB, leading to a loss of BTB integrity[3].