Introduction of a cis -Prolyl Mimic in Position 7 of the Peptide Hormone Oxytocin Does Not Result in Antagonistic Activity

Institute of Chemical Sciences and Engineering, Swiss Federal Institute of Technology, EPFL-BCH, CH-1015 Lausanne, Switzerland.
Journal of Medicinal Chemistry (Impact Factor: 5.45). 11/2005; 48(21):6553-62. DOI: 10.1021/jm049205z
Source: PubMed


New insights into the structure-activity relationship of the peptide hormone oxytocin are presented. Incorporation of the novel cis-prolyl mimic 2,2-dimethyl-1,3-thiazolidine-4-carboxylic acid (pseudoproline, PsiPro) at position 7 of the hormone yielded the analogue [Cys(Psi(Me,Me)pro)]7oxytocin (1) that showed a 92-95% induction of the cis peptide bond conformation between Cys6 and PsiPro7, as determined by one- and two-dimensional NMR spectra in water and in DMSO-d6. The impact of the dimethyl moiety regarding conformation and bioactivity was investigated by the synthesis of the corresponding dihydro compound, [Cys(Psi(H,H)pro)]7oxytocin (2). Biological tests of the uterotonic activity, the pressor activity, and the binding affinity to the rat and human oxytocin receptors were carried out. As a most significant result, no antagonistic activities were found for both the cis-constrained analogue 1 and analogue 2, suggesting that a cis conformation between residues 6 and 7 of the molecule does not result in antagonistic activity. However, the about 10-fold reduction in agonistic activity of 1 as compared to oxytocin is consistent with the reduction of the trans conformation from 90% for oxytocin to 5-8% for compound 1. Compound 1 retained a high binding affinity for the oxytocin receptor, with K(i) values of 8.0 and 1.9 nM for the rat and the human receptor, respectively. The correlation between the biological activities and the cis contents obtained from NMR analysis for compounds 1, 2, and oxytocin leads to the hypothesis that a cis/trans conformational change plays an important role in oxytocin receptor binding and activation.

4 Reads
  • Source
    • "OT and VP hormones are intensively investigated using different methods and methodologies. E.g., the structure-activity relationship of the peptide hormone oxytocin is constantly investigated using NMR experiments [20], or can be a target of numerous patents as in the case of vasopressin [21]. Both studied peptides are nonapeptides. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Eight synthetic histidine analogues of oxytocin and vasopressin are subject of investigations. The spectra of the peptides have been investigated in the terahertz band. The results are obtained in the terahertz time-domain spectroscopy arrangement.
    Optica Applicata 01/2014; 44(1). DOI:10.5277/oal40114 · 0.46 Impact Factor
  • Source
    • "The results lead however to the hypothesis that the cis/trans conformational change is playing a role in OT receptor binding and activation. There was one interesting finding and thus that in the absence and presence of magnesium ions, a considerable change was observed in the pattern of the HR protons of residues 3 and 1, indicating conformational changes that influence activity (Wittelsberger et al. 2005). Also in the case of cyclic analogues, the structural analysis revealed b-turns around residues Tyr 2 and Ile 3 , which differed from the previously discussed b-turn geometry around residues 3 and 4 that was ascribed to the conformation of OT agonists (Hruby and Lebl 1987; Oldziej et al. 1995). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Incorporation of L- or D-Tic into position 7 of oxytocin (OT) and its deamino analogue ([Mpa(1)]OT) resulted in four analogues, [L-Tic(7)]OT (1), [D-Tic(7)]OT (2), [Mpa(1),L-Tic(7)]OT (3) and [Mpa(1),D-Tic(7)]OT (4). Their biological properties were described by Fragiadaki et al. (Eur J Med Chem 42:799-806, 2007). Their NMR study (NOESY, TOCSY, (1)H-(13)C HSQC spectra) is presented here. Analogues 1, 3 and 4 showed partial agonistic activity, analogue 2 was pure antagonist, suggesting that a cis conformation between residues 6 and 7 of the molecule does not result in antagonistic activity. However, the reduction in agonistic activity of analogues 1, 3 and 4 in comparison to oxytocin is consistent with the reduction of the trans conformation form. Binding affinity for the human oxytocin receptor with IC(50) value of 130, 730, 103, and 380 nM for peptides 1, 2, 3, and 4, respectively, showed lower affinity in the case of D analogues. Deamination slightly increased the affinity. The existence of both cis and trans configurations of the Cys(6)-D-Tic(7) bond is supported by observation of two sets of cross-peaks for (1)H and (13)C nuclei for most of the residues of the peptide not only in NOESY and TOCSY but also in (1)H-(13)C HSQC spectra. The MS and HPLC indicate the presence of a single molecule/peptide, and NMR data thus suggest that this second set of peaks is due to the cis conformation.
    Amino Acids 07/2010; 39(2):539-48. DOI:10.1007/s00726-009-0470-1 · 3.29 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: A peptide-like self-immolative molecular clip is required for release of active drugs from prodrugs by endopeptidases. Upon cleavage from the carrier, this clip must collapse and release the drug rapidly. A series of aminoacyl-5,5-dimethylthiaproline (Aaa-Dmt) N-(2-(4-nitrophenyl)ethyl)amides were designed. Boc-l-aminoacyl fluorides were coupled with R-DmtOH to give Boc-l-Aaa-R-DmtOH, which were converted to the Boc-l-Aaa-R-Dmt N-(2-(4-nitrophenyl)ethyl)amides. The l,S diastereomeric series was prepared by the reaction of Boc-Aaa PFP esters with S-DmtOH. The l-Aaa-Dmt N-(2-(4-nitrophenyl)ethyl)amides were allowed to cyclise to diketopiperazines (DKPs) in aqueous buffers, expelling 2-(4-nitrophenyl)ethylamine as a model for amine-containing drugs. Reaction rates were dependant on pH. In the l,R diastereomeric series, increasing steric bulk of the Aaa side-chain (Gly, Ala, Phe, Val) led to decrease in the reaction rate. However, in the l,S series, the greatest rate of reaction was observed for the most bulky amino-acid (Val), with t½=15min at pH 8.0. The effects of steric bulk and stereochemistry are rationalised through conformational analysis (NMR and X-ray crystallography) of the starting dipeptide amides, the product diketopiperazines and key analogues. Since the dipeptides are (almost) exclusively in the cis-amide conformation, trans–cis interconversion is not relevant. The data suggest that steric interactions in the reacting conformations of the dipeptide amides, as they form the tetrahedral intermediates, are the controlling factors. Thus, l-Aaa-S-Dmt amides are shown to be excellent candidates for incorporation into the design of novel prodrugs.
    Tetrahedron 11/2006; 62(48):11245-11266. DOI:10.1016/j.tet.2006.09.009 · 2.64 Impact Factor
Show more