False positive result for a peptide drug in the gene conversion assay with Saccharomyces cerevisiae strain D7.
A battery of mutagenicity tests was performed with nafarelin, an agonist analogue of luteinizing hormone releasing hormone (LHRH) containing tryptophan (Trp) and histidine (His). Included were the Ames assay and the gene conversion assay with yeast strain D7. Both tests were negative without S9 activation, and the Ames test was negative with S9, but the yeast test was positive with S9 activation. Since the yeast test is based on conversion of cells to Trp independence, release of Trp by metabolism of the drug could account for the positive result. The test was repeated using Trp instead of the drug. The result was positive even at the lowest Trp concentration. In another experiment with the drug, amino acid analysis of the incubation mixture revealed the presence of Trp but no detectable His. Since the Ames test is based on mutation to His-independent cells, these data are completely consistent with the negative result in the Ames test and the false positive result in the yeast test. These data suggest the need for caution in interpreting the results from mutagenicity assays with peptide drugs.
DePass
L R
LR
Syntex Research, Palo Alto, California 94304.
Chan
R L
RL
Jagannath
D R
DR
Heyman
I A
IA
D000595:Amino Acid Sequence; D000818:Animals; D001711:Biotransformation; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005189:False Positive Reactions; D005785:Gene Conversion; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000633:toxicity; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D008969:Molecular Sequence Data; D009152:Mutagenicity Tests; D017274:Nafarelin; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014364:Tryptophan / Q000378:metabolism
D017274:Nafarelin; D007987:Gonadotropin-Releasing Hormone; D014364:Tryptophan