Ceramide-beta-D-glucuronide: synthesis, digestion, and suppression of early markers of colon carcinogenesis

Dietary sphingolipids inhibit chemically induced colon cancer in mice. The most likely mediators of this effect are the metabolites ceramide (Cer) and sphingosine, which induce growth arrest and apoptosis in transformed cells. Sphingolipids are digested in both the upper and the lower intestine; therefore, a more colon-specific method of delivery of sphingolipids might be useful. A Cer analogue with a D-glucuronic acid attached at the primary hydroxyl of N-palmitoyl-D-sphingosine (Cer-beta-glucuronide) was synthesized and evaluated as a substrate for Escherichia coli beta-glucuronidase and colonic digestion, as well as for suppression of early events in colon carcinogenesis in CFI mice treated with 1,2-dimethylhydrazine. Purified beta-glucuronidase (EC 3.2.1.31) and colonic segments (as a source of colonic enzymes and microflora) hydrolyzed Cer-beta-glucuronide to release Cer, as analyzed by tandem mass spectrometry. More than 75% of the Cer-beta-glucuronide was cleaved in an 8-h incubation with the colonic segments. When Cer-beta-glucuronide was administered for 4 weeks as 0.025% and 0.1% of the diet (AIN 76A) to 1,2-dimethylhydrazine-treated mice, there were significant reductions in colonic cell proliferation, as determined by in vivo BrdUrd incorporation, and in the appearance of aberrant crypt foci. The effect of dietary Cer-beta-glucuronide on aberrant crypt foci correlated significantly with the length of the colon, which suggests that Cer-beta-glucuronide was most effective when there was a larger compartment for digestion. Thus, synthetic sphingolipids that target the colon for the release of the bioactive backbones offer a promising approach to colon cancer prevention.