title: Replacement of Phe6, Phe7, and Phe11 of D-Trp8-somatostatin-14 with L-pyrazinylalanine. Predicted and observed effects on binding affinities at hSST2 and hSST4. An unexpected effect of the chirality of Trp8 on NMR spectra in methanol.
abstract: An alanine scan performed in the 1970s suggested that Phe(6) and Phe(11) are required for the binding of somatostatin (SRIF-14). Molecular modeling studies and replacement of Phe(6) and Phe(11) with a cystine bridge affording ligands with the retention of high biological activity, however, led to the alternate conclusion that Phe(6) and Phe(11) stabilize the bioactive conformation of SRIF-14. Subsequent studies revealed that Phe(11) shields Phe(6) in a "herringbone" arrangement. More recently, a report from this laboratory demonstrated that Spartan 3-21G MO calculations can be invaluable in explaining SARs in medicinal chemistry. For example, the ability of benzene and indole rings to bind the Trp(8) binding pocket for SRIF-14 and the inability of pyrazine to do so was explained through differences in electrostatic potentials. To investigate the role of Phe(6) and Phe(11) more fully, we report here the synthesis of two analogues of D-Trp(8)-SRIF in which Phe(6) and Phe(11) were replaced by the pryazinylalanine congeners, respectively. The NMR spectra in D(2)O and the K(i)s fully support the proposition that Phe(11) stabilizes the bioactive conformation through pi-bonding or aromatic edge-to-face interaction and that pyrazinylalanine(6) can be shielded by Phe(11). The data also show unexpectedly that Phe(6), via the pi-bond, interacts with the receptor, consistent with the original interpretation of the alanine scan. Heretofore it had only been known that Lys(9) interacts with an aspartate anion of the receptor. These conclusions are supported by recent studies of Lewis et al. on the effects on K(i)s of Ala(6)-SRIF-14-amide at the five receptor subtargets. We also found that pyrazinylalanine(7)-D-Trp(8)-SRIF-14 does not bind, suggesting a repulsive interaction with the receptor. Taken together, our results not only validate predictions based on Spartan 3-21G MO analysis but also provide valuable information about the nature of the receptor interaction at the molecular level. Finally, the chirality of Trp(8) was unexpectedly found to have a striking effect on NMR spectra in methanol, especially at lower temperatures.
authors.affiliation: Department of Chemistry, University of Pennsylvania, Philadelphia, 19104, USA.
authors.forename: Santhosh
authors.initials: S
authors.lastname: Neelamkavil
authors.forename: Byron
authors.initials: B
authors.lastname: Arison
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Birzin
authors.forename: Jin-Jye
authors.initials: JJ
authors.lastname: Feng
authors.forename: Kuo-Hsin
authors.initials: KH
authors.lastname: Chen
authors.forename: Atsui
authors.initials: A
authors.lastname: Lin
authors.forename: Fong-Chi
authors.initials: FC
authors.lastname: Cheng
authors.forename: Laurie
authors.initials: L
authors.lastname: Taylor
authors.forename: Edward R
authors.initials: ER
authors.lastname: Thornton
authors.forename: Amos B
authors.initials: AB
authors.lastname: Smith
authors.forename: Ralph
authors.initials: R
authors.lastname: Hirschmann
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry*; D001665:Binding Sites; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry*; D000432:Methanol; D008958:Models, Molecular; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry; D010649:Phenylalanine / Q000737:chemistry*; D011719:Pyrazines / Q000737:chemistry*; D017481:Receptors, Somatostatin / Q000737:chemistry*; D012997:Solvents; D013004:Somatostatin / Q000031:analogs & derivatives* / Q000138:chemical synthesis* / Q000737:chemistry; D013237:Stereoisomerism; D013329:Structure-Activity Relationship
chemical_list: D008565:Membrane Proteins; C501756:N-(9-fluorenylmethoxycarbonyl)pyrazinylalanine; D010456:Peptides, Cyclic; D011719:Pyrazines; D017481:Receptors, Somatostatin; D012997:Solvents; C098681:somatostatin receptor subtype-4; D010649:Phenylalanine; D013004:Somatostatin; C017783:somatostatin, Trp(8)-; C093733:somatostatin receptor 2; D000409:Alanine; D000432:Methanol