Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1999-05-06
2000-08-22
Elliott, George C.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536 253, 536 2531, 536 2532, 536 231, 435 911, C07H 2100, C07H 2102, C07H 2104
Patent
active
061074799
DESCRIPTION:
BRIEF SUMMARY
The present application is directed to a process for the preparation or purification of an oligomeric compound.
Oligomeric compounds, in particular biopolymers such as oligonucleotides, have attracted great attention because of their potential therapeutic applicability. For example, the use of oligonucleotides in antisense-technology-based therapy is subject of intensive research. Accordingly, there is a need for providing processes for the preparation or purification of oiigomeric compounds.
Oligomeric compounds, for example oligonucleotides, which structure comprises a chain of building blocks, are frequently synthesized by consecutive reaction cycles, whereby each reaction cycle comprises a step of chain elongation by reacting the growing chain of the oligomeric compound with the next chain building block. However, such a chain elongation step frequently does not go to completion so that not elongated oligomeric compounds result, which do not extend to the intended full length. In order to prevent undesired further chain elongation of such unreacted, not extended oligomeric compounds in subsequent reaction cycles, a capping step is used in each cycle to inactivate said unreacted reactive group. Furthermore, not elongated oligomeric compounds have to be separated from the final, full length oligomeric compound in the course of the preparation process.
For example, a common approach for the synthesis of oligonucleotides is the solid phase method using phosphoramidite chemistry (S. L. Beaucage and R. P. lyer, Tetrahedron 48 (1992), pp. 2223-2311), using for example acetic anhydride as capping reagent. An oligonucleotide synthesized according to this procedure usually has to be purified from undesired failure sequences. For the typical purification procedure the final terminal protecting trityl-group is not removed ("trityl-on") during oligonucleotide synthesis. Rather, the trityl-bearing full length oligonucleotide is separated from failure sequences by reverse phase HPLC. However, this method displays two main disadvantages: a) it requires a post-purification chemical treatment (removal of the trityl-group with aqueous acetic acid) and b) the relative instability of a trityl-group, like 4,4'-dimethoxytrityl (DMT), may lead to the loss of some full-length oligonucleotides due to unwanted detritylation during the purification procedure.
It is an object of the present invention to provide a process for the preparation of an oligomeric compound.
It is a further object of the present invention to provide a process for the purification of an oligomeric compound.
According to an aspect of the present invention, a process is provided for the preparation of an oligomeric compound, comprising introduction of a lipophilic capping group to an unreacted reactive group, suitable for chain elongation, of a not elongated oligomeric compound intended to be elongated in a preceeding chain-elongation step, by reacting a lipophilic capping compound with said unreacted reactive group, which lipophilic capping group is not removable under the applied conditions of the synthesis and work-up of the oligomeric compound; and which not elongated oligomeric compound capped with said lipophilic capping group can be separated from said oligomeric compound on a hydrophobic stationary phase. In this context, the term "not elongated oligomeric compound" designates an oligomeric compound not extented to its desired full length.
In a further embodiment, the process of preparation according to the present invention further comprises, after completion of the chain elongation of the oligomeric compound, full deprotection of the oligomeric compound, where a protected oligomeric compound is present. Full deprotection comprises deprotecting the terminal reactive group of the chain as well as any other protected group present in the oligomeric compound to be synthesized.
In a further embodiment, the process of preparation according to the present invention further comprises the separation of the fully deprotected oligomeric compound from the thus
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Haner Robert
Natt Fran.cedilla.ois
Elliott George C.
Epps Janet L.
McCormack Myra H.
Novartis AG
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