Computational modeling was performed to predict that a series of Cinchona alkaloids can effectively bind to the RING domain of TRAF6, leading to disruption of its native binding with the Ubc13 protein, which is critical for initiating downstream events of AKT and TAK1 activations. Both in vitro and in vivo experiments distinctly demonstrated that these Cinchona alkaloids could induce early apoptosis and reduce proliferation of cancer cells. More importantly, none of the four alkaloids appeared to influence the C-terminal domain of TRAF6 and exerted no apparent side effects on the immune system; instead, they could increase levels of TNF-α, IFN-γ, and IgG. These results support the notion that the RING domain of TRFA6 could serve as a viable anti-tumor target.

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