A new cytotoxin specific for the target cells of corticotropin-releasing factor.
To develop a cytotoxic agent selective for pituitary corticotropes, an analog of CRF, [Nle21,38,Arg36]rat (r) CRF, was conjugated to the plant toxin gelonin using glutaraldehyde as a cross-linking agent. In a two-step synthesis, glutaraldehyde was first coupled to the amino-terminal of the CRF analog. This was then reacted with gelonin to form a product, which contained 20 mol [Nle21,38,Arg36]rCRF/mol gelonin, as indicated by amino acid analysis. Efficacy and specificity of the gelonin-[Nle21,38,Arg36]rCRF conjugate were assessed in cultured rat anterior pituitary cells 3 days after a 12-h exposure to the cytotoxic conjugate, CRF, or gelonin. While pretreatment with the cytotoxic gelonin-[Nle21,38,Arg36]rCRF conjugate decreased cellular ACTH content by 74%, pretreatment with appropriate concentrations of CRF had no significant effect on cellular ACTH content. The ACTH secretory response to 2.5 nM CRF was completely eliminated by pretreatment with the gelonin-[Nle21,38,Arg36]rCRF conjugate; in contrast, pretreatment with identical doses of CRF had minimal effect. Neither pretreatment with CRF nor pretreatment with the gelonin-[Nle21,38,Arg36] rCRF conjugate had any effect on the content of LH or the LH secretory response to GnRH. These results indicate the efficacy and specificity of the conjugate of [Nle21,38,Arg36]rCRF to gelonin in eliminating CRF target cells.
Schwartz
J
J
Clayton Foundation Laboratories for Peptide Biology, Salk Institute, La Jolla, California 92037.
Penke
B
B
Rivier
J
J
Vale
W
W
D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000818:Animals; D002470:Cell Survival; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000378:metabolism; D008297:Male; D009994:Osmolar Concentration; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D014118:Toxins, Biological / Q000138:chemical synthesis* / Q000494:pharmacology
D014118:Toxins, Biological; D000324:Adrenocorticotropic Hormone; D007986:Luteinizing Hormone; D003346:Corticotropin-Releasing Hormone