Nucleotide monomers containing 8-azapurin bases or a derivative thereof, their preparation and their use in making modified olignonucleotides

• US 5,789,562 A
• Filed: 09/29/1997
• Issued: 08/04/1998
• Est. Priority Date: 05/02/1994
• Status: Expired due to Term

• Current Assignee: Sanofi-Aventis Deutschland GmbH (Sanofi )
• Original Assignee: Hoechst Aktiengesellschaft (Sanofi )
• Inventors: Seela, Frank, Lampe, Sigrid
• Primary Examiner(s): Marschel, Ardin H.
• Assistant Examiner(s): Riley, Jezia
• Application Number:
  US08/940,196
• Time in Patent Office: 309 Days
• Field of Search: 435/6, 514/1, 514/44, 536/22.1, 536/24.3, 536/27.1, 536/27.11, 536/27.13
• US Class Current: 536/22.1
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07H 19/056 Triazole or tetrazole radicals
• CPC Class Codes: C07H 19/16 Purine radicals
• CPC Class Codes: C07H 19/20 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 15/1132 against retroviridae, e.g. HIV
• CPC Class Codes: C12N 15/1133 against herpetoviridae, e.g. HSV
• CPC Class Codes: C12N 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/333 Modified A
• CPC Class Codes: C12N 2310/336 Modified G
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Y 207/11022 Cyclin-dependent kinase (2.7.11.22)
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention relates to novel modified oligonucleotides which contain at least one 8-azapurine base and form more stable hybridization complexes with nucleic acids; To a process for their preparation, and to their use as inhibitors of gene expression, as probes for detecting nucleic acids, as aids in molecular biology, and as a pharmaceutical or diagnostic agent.

Claims

1. A nucleotide monomer of the formula V ##STR19## in which V is oxy, sulfanediyl or imino;

Y^b is oxy, sulfanediyl, or methylene;

a is oxy or methylene;

R^2b is hydrogen, OR¹², C₁ -C₁₈ -alkoxy, C₁ -C₆ -alkenyloxy, halogen, azido or NR¹⁰ R¹¹ ;

R¹ is a protective group which is customary in nucleotide chemistry;

R^1b is a radical of the formula IIc or IId ##STR20## in which U is O--R⁷ or S--R⁷ ;

Q is a radical --NR⁸ R⁹ ;

R⁷ is --(CH₂)₂ --CN;

R⁸ and R⁹ are identical or different and are C₁ -C₆ -alkyl, or, together with the nitrogen atom carrying them, are a 5-9-membered heterocyclic ring which can additionally contain a further heteroatom selected from the group O, S, and N,

E'"'"' and F'"'"' are, independently of each other, H, OH, or NR¹⁰ R¹¹,

R¹⁰ and R¹¹ are identical or different and are hydrogen or an amino protective group which is customary in nucleotide chemistry, or R¹⁰ and R¹¹ together form an amino protective group which is customary in nucleotide chemistry,

R¹² is a hydroxyl protective group which is customary in nucleotide chemistry,

and the curved bracket indicates that R^2b and the adjacent phosphoryl radical can be in the 2'"'"' and 3'"'"' positions, or else, conversely, in the 3'"'"' and 2'"'"' positions.

2. A process for preparing a nucleotide monomer of the formula V as claimed in claim 1, said process comprising the steps of:

introducing onto an unprotected nucleotide monomer of the formula V suitable amino protective groups or hydroxyl protective groups, and

converting the resulting protected monomers into the corresponding phosphonate derivatives or phosphoamidite derivatives thereof.

3. A method for preparing oligonucleotides which form stable hybrids with their target nucleic acids, said method comprising condensing nucleotide monomers of formula V according to claim 1 onto an appropriately derivatized support or onto a growing oligomer chain.

5. A nucleotide monomer of formula V according to claim 1, wherein

R^2b is hydrogen, OR¹², C₁ -C₁₈ -alkoxy, allyloxy, halogen, azido or NR¹⁰ R¹¹,

R⁸ and R⁹ are isopropyl or ethyl,

R¹² is t-butyldimethylsilyl, triisopropylsilyl, o-nitrobenzyl, p-nitrobenzyl, tris(1-methylethyl)silyl or 2-fluorophenyl-4-methoxy-piperidin-4-yl.

6. A nucleotide monomer of formula V according to claim 1, wherein when R⁸ and R⁹ are a 5-9-membered heterocyclic ring, the ring additionally contains a further heteroatom selected from ##STR22##

4. A compound of the formula VI ##STR21## in which, independently of each other, U'"'"'=U"=U"'"'"' is hydroxyl or mercapto;

e and f are 0 or 1;

R¹³ is hydrogen or OH, and

E and F are H, OH or NH₂,

excluding compounds of the formula VI in which U'"'"', U", U"'"'"', R¹³ and F are OH, E is NH₂, and e and f are 1.