A simple method for the synthesis of ceramides and radiolabeled analogues

Abstract

A simple method has been developed for the coupling of amines to carboxylic acids. N-Fatty-acyl-sphingosine, cerebroside, and GM3, as well as their respective [14C] analogues, were synthesized using diethylphosphoryl cyanide as a potent coupling agent in the presence of triethylamine. The reaction procedure is rapid, racemization-free, and utilizes acids without derivatization. The desired ceramide products were obtained in 85-90% yield within one hour. The facile method presented here can also be used to synthesize [3H]-N-acyl-labeled, as well as [14C] [3H] double-labeled, ceramides.

Occurrence of ceramides and neutral glycolipids with unusual long‐chain base composition in purified rat liver mitochondria

“…However, since the borohydride treatment can also give d18:0 sphinganine by reduction of 3-ketosphingosine, the fragmentation pattern of synthetic 3ketosphingosine-containing ceramide [24] was determined and, after methanolysis and derivatisation with hepta£uorobutyric anhydride, the results with major ions at m/z 463 and 493 ruled out the presence of 3-ketosphingosine. Moreover, the analysis of synthetic ceramides made by coupling standard 3-ketosphinganine with palmitic acid [23] gave a fragmentation pattern of the long-chain base similar to that of glucosylceramide puri¢ed from rat liver mitochondria. Thus, these data con¢rm the identi¢cation as d18:0 3-ketosphinganine of the major long-chain base of glucosylceramide puri¢ed from rat liver mitochondria.…”

[14C]Ceramide Synthesis by Sphingolipid CeramideN-Deacylase: New Assay for Ceramidase Activity Detection

“…There have been several reports on the chemical synthesis of ceramide (15)(16)(17)(18). However, we wish to embe detected.…”

“…᭧ 1997 Academic Press ceramidase. To date, [ 14 C]-labeled ceramide has been prepared by purely chemical procedures (15)(16)(17)(18). This paper describes an innovative method for synthesis of [ 14 C]ceramide with high specific activity using the reCeramide, precursor of all sphingolipids, has been verse hydrolysis reaction of SCDase.…”

Chemical and enzymatic transacylation of amide‐linked FA of buttermilk gangliosides

“…The chemical FA exchange of gangliosides was established by a basic hydrolysis (29) combined with a diethylphosphorylcyanide/triethylamine-catalyzed reacylation with free arachidic acid, based on the method of Anand et al (27). The heterogeneous FA composition of native gangliosides was reduced to mainly palmitic acid and stearic acid.…”

“…The enzyme also catalyzes the condensation reaction between FFA and sphingolipids (26). Likewise, the FA exchange of gangliosides is made possible by chemical hydrolysis combined with a diethylphosphorylcyanide/triethylaminecatalyzed transacylation (27). In the biosynthesis of ganglioside, the first step in the formation of ceramide is the reduction of 3-ketosphinganine followed by the acylation of sphinganine to dihydroceramide (4).…”