Synthesis of a corticotropin analog that retains full biological activity after iodination

In order to circumvent the drastic decrease in biological activity that accompanies iodination of corticotropin, we have synthesized an analog of the human hormone containing phenylalanine in place of tyrosine in position 2 and norleucine in place of methionine in position 4 to give [Phe2,Nle4]ACTH-(1-38). This analog, referred to as Phe2,Nle4-ACTH-(1-38), was purified by ion exchange chromatography and partition chromatography. Phe2,Nle4-ACTH-(1-38) was found to be indistinguishable from synthetic human ACTH in its ability to stimulate corticosterone production in isolated rat adrenocortical cells. Iodination of Phe2,Nle4-ACTH-(1-38) resulted in the formation of monoiodo-Tyr23,Phe2,Nle4-ACTH-(1-38) and diiodo-Tyr23,Phe2,Nle4-ACTH-(1-38), which were completely separated from each other and unmodified Phe2,Nle4-ACTH-(1-38) by reverse phase high performance liquid chromatography. Monoiodo-Tyr23,Phe4,Nle4-ACTH-(1-38) was also found to be as potent as ACTH in stimulating steroidogenesis. These results show that Phe2,Nle4-ACTH-(1-38) can be used for the preparation of the radioligand with full biological activity for studying physiologically relevant corticotropin receptors and for use in RIA.