Investigating the Role of T7 and T12 Residues on the Biological Properties of Thrombin-Binding Aptamer: Enhancement of Anticoagulant Activity by a Single Nucleobase Modification

被引：46

作者：

Borbone, Nicola ^{[1
]}

Bucci, Mariarosaria ^{[2
]}

Oliviero, Giorgia ^{[1
]}

Morelli, Elena ^{[3
]}

Amato, Jussara ^{[1
]}

D'Atri, Valentina ^{[1
]}

D'Errico, Stefano ^{[1
]}

Vellecco, Valentina ^{[2
]}

Cirino, Giuseppe ^{[2
]}

Piccialli, Gennaro ^{[1
]}

Fattorusso, Caterina ^{[1
]}

Varra, Michela ^{[1
]}

Mayol, Luciano ^{[1
]}

Persico, Marco ^{[1
]}

Scuotto, Maria ^{[1
]}

机构：

[1] Univ Naples Federico II, Dipartimento Chim Sostanze Nat, I-80131 Naples, Italy

[2] Univ Naples Federico II, Dipartimento Farmacol Sperimentale, I-80131 Naples, Italy

[3] Univ Naples Federico II, Dipartimento Chim Farmaceut & Tossicol, I-80131 Naples, Italy

来源：

JOURNAL OF MEDICINAL CHEMISTRY | 2012年 / 55卷 / 23期

关键词：

G-QUADRUPLEX STRUCTURES; DNA APTAMER; INHIBITS THROMBIN; IN-VITRO; STABILITY; D(GGTTGGTGTGGTTGG); OLIGONUCLEOTIDES; PROTHROMBINASE; SUPPRESSION; RECOGNITION;

D O I：

10.1021/jm301414f

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

An acyclic pyrimidine analogue, containing a five-member cycle fused on the pyrimidine ring, was synthesized and introduced at position 7 or 12 of the 15-mer oligodeoxynucleotide GGTTGGTGTGGTTGG, known as thrombin-binding aptamer (TBA). Characterization by H-1 NMR and CD spectroscopies of the resulting aptamers, TBA-T(7)b and TBA-T(12)b, showed their ability to fold into the typical antiparallel chairlike G-quadruplex structure formed by TBA. The apparent CD melting temperatures indicated that the introduction of the acyclic residue, mainly at position 7, improves the thermal stability of resulting G-quadruplexes with respect to TBA. The anticoagulant activity of the new molecules was then valued in PT assay, and it resulted that TBA-T(7)b is more potent than TBA in prolonging clotting time. On the other hand, in purified fibrinogen assay the thrombin inhibitory activity of both modified sequences was lower than that of TBA using human enzyme, whereas the potency trend was again reversed using bovine enzyme. Obtained structure-activity relationships were investigated by structural and computational studies. Taken together, these results reveal the active role of TBA residues T-7 and T-12 and the relevance of some amino acids located in the anion binding exosite I of the protein in aptamer-thrombin interaction.

引用

页码：10716 / 10728

页数：13

共 54 条

[1] SELECTION OF SINGLE-STRANDED-DNA MOLECULES THAT BIND AND INHIBIT HUMAN THROMBIN [J].