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Synlett 2009(14): 2269-2272  
DOI: 10.1055/s-0029-1217812
LETTER
© Georg Thieme Verlag Stuttgart ˙ New York

A Novel Approach to the Preparation of Peptide-Oligonucleotide Conjugates

Renata Kaczmarek, Janina Baraniak*, Wojciech J. Stec
Department of Bioorganic Chemistry, Center of Molecular and Macromolecular Studies, Polish Academy of Sciences, Sienkiewicza 112, 90-363 Łódż, Poland
Fax: +48(42)6815483; e-Mail: baraniak@bio.cbmm.lodz.pl;
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Publication History

Received 18 May 2009
Publication Date:
07 August 2009 (online)

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Abstract

A novel approach to the synthesis of peptide-oligonucleotide conjugates (POC) based on the oxathiaphospholane chemistry has been developed. Peptide and oligonucleotide fragments, which were separately prepared, were linked postsynthetically by the functionalization of the peptide either by attachment of the oxathiaphospholane residue directly at the N-terminus or at the hydroxy group of the linker connected to N-terminus. The conjugation reaction, based on DBU-assisted nucleophilic attack of hydroxy group of oligonucleotide on phosphorus atom in the oxathiaphospholane derivatives of peptide, furnished the desired POC.

Key words

conjugation - peptides - oligonucleotides - phosphorylations - solid-phase synthesis

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16

Reaction Conditions: The synthesis of resin bound tetrathymidine phosphorothioates (4) was carried out manually by the syringe technique on 1 µmol scale and after oligonucleotide-chain assembly the coupling reaction was performed using 20-fold molar excess of 3 and 50-fold molar excess of DBU, coupling time 15 min.

17

Compound 3 underwent cyclization immediately into 5 in the presence of 1.2 equiv of DBU in CH2Cl2 solution. Data for 5: ³¹P NMR (81 MHz, CD3CN): δ = 60.70, 53.30 ppm. MS-FAB: m/z = 322 [M - 1]. Yield 79%.

18

HPLC analysis were performed using Econosil C-18 column (4.6 × 250 mm), 1.0 mL/min flow rate; buffer A, 0.1 M TEAB (pH 7.5); buffer B, 40% MeCN in 0.1 M TEAB; gradient from 0% → 60% B over 30 min, 60% over 5 min, 60% → 0% over 5 min. Yield was calculated based on A260 units of starting crude oligonucleotide and conjugate.