title: Dimeric calixarenes: a new family of major-groove binders.
abstract: A new class of potent DNA binding agents is presented. Dimeric calix[4]arenes with cationic groups at their upper rims and flexible alkyl bridges can be synthesized from triply acyl-protected calix[4]arene tetramines in relatively short synthetic sequences (3-5 steps). The compounds attach themselves to double-stranded nucleic acids in a noncovalent fashion, with micro- to nanomolar affinities. Guanidinium headgroups with their extended hydrogen-bonding "fingers" are more powerful than ammonium groups, and the benzylamine series is superior to the anilinium series (see below). The new ligands easily distinguish between RNA and various DNA types, and produce characteristic changes in UV/Vis, fluorescence, CD, as well as NMR spectra. Especially extended oligonucleotides of more than 100 base pairs are bound with affinities increasing from RNA (10 μM K(d))<AT-rich (1 μM)<GC-rich DNA double strands (100-10 nM). Ethidium bromide displacement studies confirm this order. CE(50) values are remarkably low (1-4 μM), and are more than 300 times lower than that of spermine, which is a typical backbone binder. Stoichiometries are rather high (one calixarene dimer per two BP), suggesting a potential aggregation of bound ligands inside the major groove. Most UV/Vis melting curves display an inverted shape, and start from drastically enhanced absorption intensities for the DNA complexes. DAPI displacement studies prove that up to one equivalent of calixarene dimer can be accommodated in the dye-loaded DNA. RNA complexation by calixarene dimers is accompanied by a drastic CD spectral transition from the typical A-form to a perfect B-signature, providing further experimental evidence for major-groove binding. The orientation of the ligands can be deduced from NMR titrations and is reproduced in Monte-Carlo simulations on 1:1 complexes in water.
authors.affiliation: Department of Chemistry, Universität Duisburg-Essen, Universitätsstr. 7, 45117 Essen, Germany.
authors.forename: Wenbin
authors.initials: W
authors.lastname: Hu
authors.forename: Caroline
authors.initials: C
authors.lastname: Blecking
authors.forename: Marijeta
authors.initials: M
authors.lastname: Kralj
authors.forename: Lidija
authors.initials: L
authors.lastname: Šuman
authors.forename: Ivo
authors.initials: I
authors.lastname: Piantanida
authors.forename: Thomas
authors.initials: T
authors.lastname: Schrader
mesh_terms: D020029:Base Pairing; D001665:Binding Sites; D047250:Calixarenes / Q000737:chemistry*; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D006860:Hydrogen Bonding; D008024:Ligands; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D008024:Ligands; D009841:Oligonucleotides; D047250:Calixarenes; D004247:DNA