Background: Chronic myeloid leukemia (CML) is currently treated with imatinib, a Bcr-Abl
inhibitor. However, resistance to this drug usually develops over time. Triptolide, a diterpenoid triepoxide, has
been shown active against CML cells resistant to imatinib, acting mainly on the level of Bcr-Abl transcription
Objective: Here, we used the triterpene betulinic acid, a known proteasome inhibitor with potential antileukemic
activity, as a scaffold for the generation of analogues with predicted triptolide biological activity.
Method: Betulinic acid derivatives were designed based on the structure-activity relationship of triptolide and
evaluated for their cytotoxic effects in CML cells, lymphocytes and human keratinocytes (HaCaT), as well as
against the proteasome complex. The main modification performed on betulinic acid was fluorination at C-28
and epoxidation, both of which are responsible for enhancing activity of triptolide. A total of 10 compounds
were obtained: 6 previously described and 4 novel compounds. The cytotoxic activity over a CML cell line
(K562) was assessed using flow cytometry and compared to lymphocytes and HaCaT.
Result: The results show that betulinic acid was the most cytotoxic compound against CML cells, showing a
good selectivity index for cancer over normal cells. The most important trend for the activity in betulinic
acid derivatives is the presence of a free hydroxyl group at C-3 and a carboxyl group at C-28. Results also
indicated that the epoxide is important for enhancing the activity, while modification at C-28 worsens the
Conclusion: Proteasome inhibition assays suggest that proteasome is the main target for betulinic acid and its