Synthesis of selenium-derivatized nucleosides, nucleotides, phosphoramidites, triphosphates and nucleic acids

• US 8,354,524 B2
• Filed: 06/29/2011
• Issued: 01/15/2013
• Est. Priority Date: 03/22/2001
• Status: Active Grant

• Current Assignee: Georgia State University Research Foundation, Inc.
• Original Assignee: Georgia State University Research Foundation, Inc.
• Inventors: Huang, Zhen
• Primary Examiner(s): Balasubramanian, Venkataraman
• Application Number:
  US13/172,559
• Publication Number:
  US 20120065387A1
• Time in Patent Office: 566 Days
• Field of Search: 544/317, 536/28.1, 536/26.8
• US Class Current: 536/26.8
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• 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: C07H 5/08 to sulfur, selenium or tellurium

Abstract

The present invention provides selenium derivatives of nucleosides, nucleoside phosphoramidites, nucleotides, nucleotide triphosphates, oligonucleotides, polynucleotides, and larger nucleic acids and methods for their synthesis. Selenium derivatives of both ribonucleic acids and deoxyribonucleic acids, as well as methods for their synthesis, crystallization and uses in structural determinations, particularly by X-ray crystallographic techniques are disclosed. The selenium derivatives of the present invention are also useful as food supplements.

Claims

1. A selenium-containing compound having the structure of formula I:

2. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is a nucleic acid base selected from the group consisting of cytosine and 2-selenocytosine;

or is a protected nucleic acid base selected from the group consisting of protected cytosine and 2-selenocytosine;

(b) R₂ is H, HO, alkyl-O, TBDMS-O, TOM-O group, ACE-O group, methyl-Se, ethyl-Se, or propyl-Se;

(c) R₃ is H, HO, alkyl-O, TBDMS-O, methyl-Se, ethyl-Se, propyl-Se, phosphoramidite, or phosphate;

(d) R₄ is H, HO, alkyl-O, DMTr-O, methyl-Se, ethyl-Se, propyl-Se, phosphate, diphosphate, diphosphoroselenoate, triphosphate, or triphosphoroselenoate; and

(e) X is an oxygen atom;

wherein at least one of R₁ or R₂ comprises at least one selenium atom.

3. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is a nucleic acid base selected from the group consisting of cytosine and 2-selenocytosine;

or is a protected nucleic acid base selected from the group consisting of protected cytosine and 2-selenocytosine;

(b) R₂ is H, alkyl-O, TBDMS-O, TOM-O group, ACE-O group, methyl-Se, ethyl-Se, or propyl-Se;

(c) R₃ is phosphoramidite;

(d) R₄ is DMTr-O; and

(e) X is an oxygen atom;

wherein at least one of R₁ or R₂ comprises at least one selenium atom.

4. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is the nucleic acid base cytosine;

or is a protected cytosine;

(b) R₂ is methyl-Se;

(c) R₃ is phosphoramidite;

(d) R₄ is DMTr-O, TBDMS-O, or (alkyl-O)₃Si—

O; and

(e) X is an oxygen atom.

5. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is a protected 2-selenocytosine;

(b) R₂ is H, alkyl-O, TBDMS-O, TOM-O group, or ACE-O group;

(c) R₃ is phosphoramidite;

(d) R₄ is DMTr-O; and

(e) X is an oxygen atom.

6. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is the nucleic acid base cytosine;

(b) R₂ is diselenide, or alkyl-Se;

(c) R₃ is HO or H;

(d) R₄ is HO or H; and

(e) X is an oxygen atom.

7. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is 2-selenocytosine;

(b) R₂ is HO or H;

(c) R₃ is HO or H;

(d) R₄ is HO or H; and

(e) X is an oxygen atom.

8. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is the nucleic acid base cytosine;

(b) R₂ is HO or H;

(c) R₃ is HO or H;

(d) R₄ is HO or H; and

(e) X is a selenium atom.

9. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is a nucleic acid base selected from cytosine and 2-selenocytosine;

(b) R₂ is H, HO, or methyl-Se;

(c) R₃ is H, HO, or methyl-Se;

(d) R₄ is phosphate, diphosphate, diphosphoroselenoate, triphosphate, or triphosphoroselenoate; and

(e) X is an oxygen atom;

wherein at least one of R₁ or R₂ comprises at least one selenium atom.

10. A selenium-containing compound according to claim 1, wherein:

(a) R₁ is 2-selenocytosine;

(b) R₂ is H or HO;

(c) R₃ is HO;

(d) R₄ is triphosphate; and

(e) X is an oxygen atom.

11. A process for preparing a compound having the structure of formula I:

12. A process according to claim 11, wherein:

(a) R₁ is a nucleic acid base selected from the group consisting of cytosine and 2-selenocytosine;

or is a protected nucleic acid base selected from the group consisting of protected cytosine and 2-selenocytosine;

(b) R₂ is H, HO, alkyl-O, TBDMS-O, TOM-O group, ACE-O group, methyl-Se, ethyl-Se, or propyl-Se;

(c) R₃ is H, HO, alkyl-O, TBDMS-O, methyl-Se, ethyl-Se, propyl-Se, phosphoramidite, or phosphate;

(d) R₄ is H, HO, alkyl-O, DMTr-O, methyl-Se, ethyl-Se, propyl-Se, phosphate, diphosphate, diphosphoroselenoate, triphosphate, or triphosphoroselenoate; and

(e) X is an oxygen atom;

wherein at least one of R₁, or R₂ comprises at least one selenium atom.

13. A process according to claim 11, wherein:

(a) R₁ is a nucleic acid base selected from the group consisting of cytosine and 2-selenocytosine;

or is a protected nucleic acid base selected from the group consisting of protected cytosine and 2-selenocytosine;

(b) R₂ is H, alkyl-O, TBDMS-O, TOM-O group, ACE-O group, methyl-Se, ethyl-Se, or propyl-Se;

(c) R₃ is phosphoramidite;

(d) R₄ is DMTr-O; and

(e) X is an oxygen atom;

wherein at least one of R₁ or R₂ comprises at least one selenium atom.

14. A process according to claim 11, wherein the water immiscible organic phase comprises toluene, benzene, or hexane.

15. A process according to claim 11, wherein the phase transfer catalyst comprises a quaternary ammonium ion and a counterion.

16. A process according to claim 11, wherein the phase transfer catalyst comprises a quaternary ammonium ion and a counterion selected from the group consisting of F⁻

, Cl⁻

, Br⁻

, I⁻

ClO₃⁻

, NO₃⁻

, HCO₃⁻

, and HSO₄⁻

.

17. A process according to claim 11, wherein an organic solvent for one phase reaction is ethanol, DMF, acetonitrile, or a mix of solvents.