Abstract
The 13-residue peptide, χ-conotoxin MrIA extracted from the venom of Conus marmoreus, is a potent and selective inhibitor of the human noradrenaline transporter (NET). With the aim of improving its biophysical properties, chemical modifications were performed including the attachment of a lipophilic amino acid at the N-terminus and cyclisation of the peptide backbone with functionality introduced into the linker. All χ-conotoxin MrIA analogues were assembled on solid phase by highly optimised Boc chemistry and N- to C-cyclic analogues accessed by cysteine-mediated intramolecular native chemical ligation. In vitro biological activity at the human NET was evaluated by functional assays. All analogues inhibited the uptake of [3H]noradrenaline with comparable potencies to that of the native peptide, with one of the analogues, the linear N-terminal aminotetradecanoyl MrIA showing a 3-fold increase in potency (p < 0.05).
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Abbreviations
- Ac:
-
acetyl
- Atda:
-
(D,L)-2-aminotetradecanoic acid
- Boc:
-
tert-butyloxycarbonyl
- BrZ:
-
2-bromobenzyloxycarbonyl
- BSA:
-
bovine serum albumin
- Bzl:
-
benzyl
- ClZ:
-
2-chlorobenzyloxycarbonyl
- DIEA:
-
diisopropylethylamine
- DMEM:
-
Dulbecco's Modified Eagle's Medium
- DMF:
-
N,N-dimethylformamide
- DMSO:
-
dimethylsulfoxide
- Dnp:
-
dinitrophenyl
- GnHCl:
-
guanidine hydrochloride
- HBTU:
-
N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminium hexafluorophosphate N-oxide
- HF:
-
anhydrous hydrogen fluoride
- Hyp (O):
-
4-hydroxyproline
- MeBzl:
-
4-methylbenzyl
- MESNA:
-
sodium 2-mercaptoethanesulfonate
- NET:
-
noradrenaline transporter
- PAM:
-
phenylacetamidomethyl
- RP-HPLC:
-
reversed-phase high performance liquid chromatography
- TFA:
-
trifluoroacetic acid
- TIPS:
-
triisopropylsilane
- Tos:
-
toluene sulfonyl
- Xan:
-
xanthenyl
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Dekan, Z., Paczkowski, F.A., Lewis, R.J. et al. Synthesis and In vitro Biological Activity of Cyclic Lipophilic χ-Conotoxin MrIA Analogues. Int J Pept Res Ther 13, 307–312 (2007). https://doi.org/10.1007/s10989-007-9083-2
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DOI: https://doi.org/10.1007/s10989-007-9083-2