Dr. Mills first evaluated the activity of COTI-2 in a number of human cancer cells lines that we had not previously studied. He was able to evaluate the killing activity of COTI-2 in the context of p53 mutations since the p53 mutation status of these cells was known. He then went on to introduce 32 different types of p53 mutations into mouse embryonic fibroblasts (MEFs), a cell-based system for studying the effect of gene mutations. Once the p53 gene mutations were established in the MEFs the effect of COTI-2 on those cells was evaluated.
Changes in normal p53 protein levels together with changes in other proteins known to be direct cellular targets of normal p53 protein were also evaluated for each of 32 different types of p53 mutations following treatment with COTI-2.
Extensive data from the experiments have demonstrated that COTI‐2 restores p53 protein to its normal function in human cancer cells with a wide range of common p53 gene mutations.