Previous investigations from our laboratory have demonstrated a significant reduction in the catalytic function of the 60 kDa and 63 kDa isozymes of calmodulin-dependent cyclic nucleotide phosphodiesterase (CaMPDE) when comparing human cerebral tissue that was free of tumor and glioblastoma multiforme (GBM) and gliosarcoma [Lal S., Raju R. V. S., Macaulay R. B. J., and Sharma R. K. (1996) Can. J. Neurol. Sci., 23, 245–250], The results suggested the possibility of an endogenously produced inhibitor of CaMPDE expressed in these tumors. Further investigation has initially characterized the presence of a heat-labile, protein inhibitor of both the 60 kDa and 63 kDa isozymes of CaMPDE. Sephacryl S-200 gel filtration column chromatography indicated that the inhibitor has an apparent molecular weight of 22 kDa and experimental evidence demonstrates that this inhibitor protein may act independently of calmodulin, and is therefore a novel CaMPDE inhibitor. Previous work on non-CNS tumors has shown high levels of CaMPDE activity and absence of an inhibitor. This suggests that a different mechanism may exist for the proliferation of these subsets of tumors.