Increase in melanin formation and promotion of cytotoxicity in cultured melanoma cells caused by phosphorylated isomers of L-dopa.
A new class of compounds, termed "dopa phosphates," is described. The compounds contain phosphate ester linkages at positions 3 and/or 4 of the phenylalanine ring. Dopa phosphates are highly soluble compounds which are stable over a wide range of pH values and are not hydrolyzed by boiling in concentrated acid. Synthetic yields of greater than 90% can be obtained using dopa as starting material. Exposure to alkaline phosphatase results in hydrolysis of the phosphate moieties and production of dopa. Dopa phosphates do not inhibit dopa oxidase (tyrosinase, EC 1.14.18.1) activity. Dopa oxidase does not catalyze the conversion of dopa phosphates into melanin unless the dopa phosphates are first treated with alkaline phosphatase. Dopa phosphates, when compared to L-dopa, are stable in the presence of O2 and are not oxidized by serum proteins. In the presence of cultured melanoma cells, dopa phosphates are readily converted into melanin, indicating that the cells are able to produce dopa from dopa phosphates. At high concentrations, dopa phosphates are cytotoxic toward melanoma cells in culture. The cytotoxicity is enhanced at least 3-fold by pretreatment of cells with melanotropin and is prevented by phenylthiourea, an inhibitor of dopa oxidase activity. These results, combined with studies on the uptake of radioactive forms of dopa phosphates (32P and 14C), indicate that phosphorylated isomers of dopa are efficiently taken up by Cloudman melanoma cells and are readily converted by the cells into a melanin precursor, presumably L-dopa.
Pawelek
J M
JM
Murray
M
M
D000818:Animals; D001692:Biological Transport; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D007980:Levodopa / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism / Q000627:therapeutic use; D008543:Melanins / Q000096:biosynthesis*; D008545:Melanoma / Q000188:drug therapy / Q000378:metabolism*; D051379:Mice; D014442:Monophenol Monooxygenase / Q000378:metabolism; D010766:Phosphorylation; D012995:Solubility
D008543:Melanins; D007980:Levodopa; D014442:Monophenol Monooxygenase