Roche Innovation Center Copenhagen A/s

Hoffmann-la Roche Inc.

F. Hoffmann-la Roche Ag

Dennis Jul Hansen

Bernd Kuhn

Luca Claudio Gobbi

Katrin Groebke Zbinden

Erik Daa Funder

Christoph Rosenbohm

Michael Meldgaard

Konrad H Bleicher

Lykke Pedersen

Jacob Ravn

Peter Hagedorn

Buelent Kocer

Carsten Kroll

Søren Vestergaard Rasmussen

Martin Ritter

Heidi Rye Hudlebusch

Uwe Grether

Hans Richter

Marianne B Mogensen

Joerg Hoernschemeyer

Rui Chen

Benoit Hornsperger

Joerg Benz

Mette Ladefoged

Fionn O`hara

Charles Bell

Satoshi Tsuchiya

Marianne Mogensen

The present invention relates to a novel process for preparing a compound of formula (III) and rhenium chelated MAG3 oligonucleotides.

Process for preparing rhenium chelated MAG3 oligonucleotides

The present invention relates to methods of synthesis of stereodefined oligonucleotides wherein the protected 5'-hydroxy terminus of a nucleoside or oligonucleotide attached to a solid support is first unprotected, followed by coupling an oxazaphospholidine phosphoramidite monomer to the now deprotected 5&#x2032;-hydroxy terminus, oxidizing the resultant phosphite triester intermediate with a sulfurizing reagent, and optionally washing the product. The method of synthesis of the present invention described above may be repeated to optionally elongate the stereodefined oligonucleotide being synthesized until the elongation cycle is intentionally terminated via deprotecting and cleavaging the resulting stereodefined oligonucleotide from the solid support. The oxazaphospholidine phosphoramidite monomer to be coupled in the present invention may optionally be chosen from Formula 1a and 1b. Generally, the present invention relates to enhanced synthesis methods wherein a single elongation cycle, optionally comprising repeated coupling and oxidation steps, results in an enhanced yield and higher purity of stereodefined phosphorothioate oligonucleotides.

Multiple coupling and oxidation method

The present invention relates to a method for preparing stereodefined phosphorothioate oligonucleotides, especially locked stereodefined phosphorothioate oligonucleotides with a high yield, using pyridinium acidic salts as a coupling activator.

Pyridinium salts as activators in the synthesis of stereodefined oligonucleotides

Recent advancements in LNA oligonucleotides include the use of amine linkers to link an LNA antisense oligonucleotide to a conjugate group. For example please see WO2014/I18267. The present invention originates from the identification of a problem when de-protecting LNA oligonucleotides which comprise an aliphatic amine group and DMF protected LNA G nucleoside, which results in the production of a +28 Da impurity. This problem is solved by using acyl protection groups on the exocyclic nitrogen of the LNA-G residue, rather than the standard DMF protection group.

LNA-G process

The present invention relates to the field of stereodefined phosphorothioate oligonucleotides and to stereodefining nucleoside monomers comprising an amine containing chiral auxiliary group, and methods of orthogonally protecting the nitrogen of chiral auxiliaries during oligonucleotide synthesis, preventing post elongation chain cleavage, increasing yield and purity of stereodefined phosphorothioate oligonucleotides.

Orthogonal protecting groups for the preparation of stereodefined phosphorothioate oligonucleotides

The present invention relates to the field of oligonucleotide conjugates and to methods of synthesis thereof. In the present method a low-water content solvent environment allows a more efficient conjugation, reducing the amount of conjugate moiety needed and increasing the conjugation reaction speed.

Oligonucleotide conjugation process

Methods for cleaving oligonucleotides from a solid support are described as are methods for synthesizing an oligonucleotide on a solid support and subsequently cleaving the oligonucleotide from the solid support. In the methods, the 3' nucleoside of the oligonucleotide attached to the solid support is a LNA nucleoside. The method entail treating the bound oligonucleotide with a concentrated ammonium hydroxide solution for about 30 minutes to about 6 hours.

Rapid unylinker cleavage

The present invention relates to antisense oligonucleotides that are capable of modulating expression of ATXN2 in a target cell. The oligonucleotides hybridize to ATXN2 mRNA. The present invention further relates to conjugates of the oligonucleotide and pharmaceutical compositions and methods for treatment of neurodegenerative diseases such as spinocerebellar ataxia type 2 (SCA2), amyotrophic lateral sclerosis (ALS), Alzheimer's frontotemporal dementia (FTD), parkinsonism and conditions with TDP-43 proteinopathies using the oligonucleotide.

Oligonucleotides for modulating ATXN2 expression

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Hørsholm, Denmark

Dennis Jul Hansen