Constrained Corticotropin-Releasing Factor (CRF) Agonists and Antagonists with i−(i+3) Glu-Xaa-d Xbb-Lys Bridges
| Autor: | J. Gulyas, Anne Z. Corrigan, Jean Rivier, S L Lahrichi, Wylie Vale, A. G. Craig, Steven C. Koerber, Catherine Rivier |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Male
Models Molecular Agonist Circular dichroism Molecular model Corticotropin-Releasing Hormone medicine.drug_class Stereochemistry Glutamine Molecular Sequence Data Peptides Cyclic Protein Structure Secondary Rats Sprague-Dawley chemistry.chemical_compound Protein structure Adrenocorticotropic Hormone Pituitary Gland Anterior Drug Discovery medicine Peptide synthesis Animals Humans Amino Acid Sequence Cells Cultured Urocortin chemistry.chemical_classification Chemistry Circular Dichroism Lysine Adrenalectomy Peptide Fragments Cyclic peptide Rats Lactam Molecular Medicine |
| Zdroj: | Journal of Medicinal Chemistry. 41:5002-5011 |
| ISSN: | 1520-4804 0022-2623 |
| DOI: | 10.1021/jm980350k |
| Popis: | We hypothesized that covalent constraints such as side-chain to side-chain lactam rings would stabilize an alpha-helical conformation shown to be important for the recognition and binding of the human corticotropin-releasing factor (hCRF) C-terminal 33 residues to CRF receptors. These studies led to the discovery of cyclo(20-23)[DPhe12,Glu20,Lys23,Nle21,38]hCRF (12-41) and of astressin ¿cyclo(30-33)[DPhe12,Nle21,38,Glu30,Lys33]hCR F(12-41)¿, two potent CRF antagonists, and of cyclo(30-33)[Ac-Leu8,DPhe12,Nle21, Glu30,Lys33,Nle38]hCRF(8-41), the shortest sequence equipotent to CRF reported to date (Rivier et al. J. Med. Chem. 1998, 41, 2614-2620 and references therein). To test the hypothesis that the Glu20-Lys23 and Glu30-Lys33 lactam rings were favoring an alpha-helical conformation rather than a turn, we introduced a D-amino acid at positions 22, 31, and 32 in the respective rings. Whereas the introduction of a D-residue at position 31 was only marginally deleterious to potency (ca. 2-fold decrease in potency), introduction of a D-residue at position 22 and/or 32 was favorable (up to 2-fold increase in potency) in most of the cyclic hCRF, alpha-helical CRF, urotensin, and urocortin agonists and antagonists that were tested and was also favorable in linear agonists but not in linear antagonists; this suggested a unique and stabilizing role for the lactam ring. Introduction of a [DHis32] (6) or acetylation of the N-terminus (7) of astressin had a minor deleterious or a favorable influence, respectively, on duration of action. In the absence of structural data on these analogues, we conducted molecular modeling on an Ac-Ala13-NH2 scaffold in order to quantify the structural influence of specific L- and DAla6 and L- and DAla7 substitutions in [Glu5,Lys8]Ac-Ala13-NH2 in a standard alpha-helical configuration. Models of the general form [Glu5,LAla6 or DAla6,LAla7 or DAla7,Lys8]Ac-Ala13-NH2 were subjected to high-temperature molecular dynamics followed by annealing dynamics and minimization in a conformational search. A gentle restraint was applied to the 0-4, 1-5, and 8-12 O-H hydrogen bond donor-acceptor pairs to maintain alpha-helical features at the N- and C-termini. From these studies we derived a model in which the helical N- and C-termini of hCRF form a helix-turn-helix motif around a turn centered at residue 31. Such a turn brings Gln26 in close enough proximity to Lys36 to suggest introduction of a bridge between them. We synthesized dicyclo(26-36,30-33)[DPhe12,Nle21,Cys26,Glu30 ,Lys33,Cys36, Nle38]Ac-hCRF(9-41) which showed significant alpha-helical content using circular dichroism (CD) and had low, but measurable potency ¿0. 3% that of 6 or ca. 25% that of [DPhe12,Nle21,38]hCRF(12-41)¿. Since the 26-36 disulfide bridge is incompatible with a continuous alpha-helix, the postulate of a turn starting at residue 31 will need to be further documented. |
| Databáze: | OpenAIRE |
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