Phosphoramidite derivatives, their preparation and the use thereof in the incorporation of reporter groups on synthetic oligonucleotides
AssigneeAmersham International plc
InventorsKonrad Misiura, Michael Gait
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Abstract
Phosphoramidite derivatives of formula (V), ##STR1## wherein X is biotin and Y is a protecting group. There may be a linker arm, of variable length, between X and the rest of the molecule. Examples of the protecting group Y include 4,4'-dimethoxytrityl, trifluoroacetyl and fluorenylmethoxycarbonyl (Fmoc). The phosphoramidite derivatives are useful in single or multiple labelling of synthetic oligonucleotides. Process for the preparation of these phosphoramidite derivatives are also disclosed.
Claims6
A phosphoramidite derivative of the following formula:
- ##STR15## wherein X=biotin, and
- Y=a protecting group.
A method for the production of a phosphoramidite derivative of the following formula:
- ##STR16## and which comprises:
- i) Reaction of solketal with acrylonitrile to form the addition product, 2-cyanoethyl solketal (I):
- ##STR17## ii) Reduction of the resultant compound (I) to form 3-aminopropyl solketal (II):
- ##STR18## iii) Reaction of the resultant compound (II) with biotin N-hydroxysuccinimide to form N-biotinyl-3-aminopropyl solketal (III):
- ##STR19## iv) Reaction of the resultant compound (III) with 4,4'-dimethoxytrityl chloride to form 1-0-(4, 4'-dimethoxytrityl)-3-0-(N-biotinyl-3-aminopropyl) glycerol (IV):
- ##STR20## v) Phosphitylation of the resultant compound (IV) to produce the desired biotinyl phosphoramidite derivative (V):
- ##STR21## wherein DMTr=4,4'-dimethoxytrityl.
Dependencies are inferred from the claim text.