MODIFIED NUCLEOTIDES

• US 20120095201A1
• Filed: 10/25/2011
• Published: 04/19/2012
• Est. Priority Date: 12/23/2002
• Status: Active Grant

• Current Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Original Assignee: Illumina Cambridge Limited (Illumina Incorporated)
• Inventors: Wu, Xiaolin, Smith, Mark, Brennan, Joseph, Barnes, Colin, Liu, Xiaohai, Ruediger, Silke, MILTON, John
• Granted Patent: US 8,597,881 B2
• Time in Patent Office: Days
• US Class Current: 536/25.32
• CPC Class Codes: C07H 1/00 Processes for the preparation of sugar derivatives
• CPC Class Codes: C07H 19/00 Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
• 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 19/14 Pyrrolo-pyrimidine 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: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2525/186 incorporating a non-extendable or blocking moiety
• CPC Class Codes: C12Q 2525/197 incorporating a spacer/coupling moiety
• CPC Class Codes: C12Q 2535/113 Cycle sequencing
• CPC Class Codes: Y02P 20/55 Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

Provided herein is a method for replacing the allyl group in a compound of formula R—O-allyl, R₂N(allyl), RNH(allyl), RN(allyl)₂ or R—S-allyl with a hydrogen, using a transition metal and one or more phosphine ligands.

Claims

1-15. -15. (canceled)

16. A method of converting a compound of formula R—

O-allyl, R₂N(allyl), RNH(allyl), RN(allyl)₂ or R—

S-allyl to a corresponding compound in which the allyl group is removed and replaced by hydrogen, said method comprising the steps of reacting a compound of formula R—

O-allyl, R₂N(allyl), RNH(allyl), RN(allyl)₂ or R—

S-allyl in aqueous solution with a transition metal comprising a transition metal and one or more ligands selected from the group comprising water-soluble phosphine and water-soluble nitrogen-containing phosphine ligands, wherein the or each R is a water-soluble biological molecule.

17. The method of claim 16 wherein said compound is of formula R—

O-allyl.

18. The method of claim 16 wherein said R is part of a nucleoside, a nucleotide or a polynucleotide molecule.

19. The method of claim 18 wherein said nucleoside, nucleotide or polynucleotide further comprises a detectable label linked to the base thereof by a cleavable or non-cleavable linker.

20. The method of claim 19 wherein said linker is cleavable.

21. The method of claim 19, wherein said detectable label is a fluorophore.

22. The method of claim 19 wherein said linker is acid labile, photolabile or contains a disulfide linkage.

23. The method of claim 19 wherein said allyl group and said label are removed in a single step.

24. The method of claim 16 wherein said transition metal is selected from the group comprising platinum, palladium, rhodium, ruthenium, osmium and iridium.

25. The method of claim 16 wherein said transition metal is palladium.

26. The method of claim 16 wherein said group of ligands comprise derivatised triaryl phosphine ligands or derivatised trialkyl phosphine ligands.

27. The method of claim 16 wherein said group of ligands are derivatised with one or more functionalities selected from the group comprising amino, hydroxyl, carboxyl and sulfonate groups.

28. The method of claim 16 wherein the group of ligands comprises 3,3′

,3″

-phosphinidynetris (benzenesulfonic acid) and tris(2-carboxyethyl)phosphines and their salts.

29-50. -50. (canceled)