Alkynylamino-nucleotides
AssigneeE. I. Du Pont de Nemours and Company
InventorsFrank W. Hobbs, Jr., Anthony J. Cocuzza
Life of this patent
Estimating the term…
Abstract
Alkynylamino-nucleotides and labeled alkynylamino-nucleotides useful, for example, as chain terminating substrates for DNA sequencing are provided along with several key intermediates and processes for their preparation.
Claims12
An alkynylamino-nucleotide having the structure:
- Nuc--C.tbd.C--R.sub.1 --NR.sub.2 R.sub.3
- wherein
- R.sub.1 is a diradical moiety of 1-20 atoms,
- R.sub.2 and R.sub.3 are, independently, H, C.sub.1 -C.sub.4 alkyl or a protecting group, and Nuc is R.sub.4 -Het having the structure ##STR16## Z is H or NH.sub.2, and R.sub.4 is ##STR17## wherein R.sub.5 is H, PO.sub.3 H.sub.2, P.sub.2 O.sub.6 H.sub.3, P.sub.3 O.sub.9 H.sub.4 or salts thereof, and
- (i) when R.sub.7 .dbd.R.sub.8 .dbd.H, then R.sub.6 .dbd.H, OH, F, N.sub.3 or NH.sub.2 ;
- or
- (ii) when R.sub.7 .dbd.H and R.sub.8 .dbd.OH, then R.sub.6 .dbd.H or OH;
- or
- (iii) when R.sub.7 .dbd.OH and R.sub.8 .dbd.H, then R.sub.6 .dbd.OH.
A labeled alkynylamino-nucleotide having the structure:
- Nuc--C.tbd.C--R.sub.1 --NR.sub.2 R.sub.3
- wherein
- R.sub.1 is a diradical moiety of 1-20 atoms.
- R.sub.2 is H, C.sub.1 -C.sub.4 alkyl or a
- protecting group, and
- Nuc is R.sub.4 -Het having the structure ##STR18## Z is H or NH.sub.2, and R.sub.4 is ##STR19## wherein R.sub.5 is H, PO.sub.3 H.sub.2, P.sub.2 O.sub.6 H.sub.3, P.sub.3 O.sub.9 H.sub.4 or
- (i) when R.sub.7 .dbd.R.sub.8 .dbd.H, then R.sub.6 .dbd.H, OH, R, N.sub.3 or NH.sub.2 ;
- or
- (ii) when R.sub.7 .dbd.H and R.sub.8 .dbd.OH, then R.sub.6 .dbd.H or OH;
- or
- (iii) when R.sub.7 .dbd.OH and R.sub.8 .dbd.H, then R.sub.6 .dbd.OH,
- and R.sub.R.sub.3 is a reporter group.
A process for preparing 2-thioalkoxy-6-alkoxy-7-iodo-7-deazapurines wherein 2-thioalkoxy-7-deazapurines are reacted with N-iodosuccinimide for the regioselective introduction of iodine at the 7-position and obtaining the 7-iodo regioisomer.
A process of preparing 7-(3-amino-1-propynyl)2',3'-dideoxyguanosine 5'-triphosphate comprising the steps of:
- (A) reacting 6-methoxy-2-methylthio-7-deazapurine with 1-chloro-2-deoxy-3,5-di-O-p-toluoyl-.alpha.-D-ribofuranose in presence of sodium hydride;
- (B) hydrolyzing the ester product 9 of step (A) under basic condition to diol 10;
- (C) protecting the 5-OH position in diol 10 with a trityl group;
- (D) removing the 3-OH group by reducing an intermediate thionocarbonate with a tin hydride reducing agent;
- (E) treating the dideoxydeazapurine 12 resulting from step (D) with N-iodosuccinimide to form the 7-iodo derivative 13;
- (F) reacting compound 13 with sodium thiocresolate in hexamethylphosphoramide to form the 7-deazapurin-6-one 14;
- (G) treating compound 14 successively with meta-chloroperoxybenzoic acid and ammonia to obtain the 7-deazaguanosine 15;
- (H) coupling N-propargyltrifluoroacetamide to the deprotected 7-iodo compound 16 to obtain 7-(3-trifluoroacetamido-1-propynyl)-2',3'-dideoxy-7-deazaguanosine;
- and
- (I) reacting the product of step (H) sequentially with phosphorous oxychloride and pyrophosphate to convert it to the 5'-triphosphate followed by deprotection.
A process for preparing 6-chloro-7-iodo-7-deazapurines wherein 6-chloro-7-deazapurines are reacted with iodine monochloride for the regioselective introduction of iodine at the 7-position and obtaining the 7-iodo regioisomer.
An improved method for preparing alkynylaminonucleotides by coupling iodo-nucleotides having an iodine atom attached to a carbon atom within the heterocyclic base with terminal alkynyl amines using a bis(triphenylphosphine)palladium dichloride/cuprous iodide catalyst, the improvement comprising the use of a tetrakis(triphenylphosphine)palladium(O)/cuprous iodide catalyst.
7-Iodo-nucleotides having the structure:
- ##STR20## Z is H or NH.sub.2, and R.sub.4 is ##STR21## wherein R.sub.5 is H, PO.sub.3 H.sub.2, P.sub.2 O.sub.6 H.sub.3, P.sub.3 O.sub.9 H.sub.4 or salts thereof, and
- (i) when R.sub.7 .dbd.R.sub.8 .dbd.H, then R.sub.6 .dbd.H, OH, R, N.sub.3 or NH.sub.2 ;
- or
- (ii) when R.sub.7 .dbd.H and R.sub.8 .dbd.OH, then R.sub.6 .dbd.H or OH;
- or
- (iii) when R.sub. .dbd.OH and R.sub.8 .dbd.H, then R.sub.6 .dbd.OH.
Dependencies are inferred from the claim text.