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US4404368A
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Lyophilized phosphorylated nucleosides
Assignee: Ens Bio Logicals Inc
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Filing: 1981-02-17
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Granted: 1983-09-13
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Expires: 2001-02-17
Abstract
Abstract Phosphorylated nucleotides for addition to nucleotide chains in the stepwise synthesis of oligonucleotides, e.g. as a polymeric support, are prepared in dioxane solvent and then lyophilized, to provide a stable, dry product ready for re-dissolving in reaction solvent. The lyophilized products are stable indefinitely and do not suffer from decreased reactivity.
Inventors
- 👤 Gabriel G. Alvarado-Urbina
- 👤 Kelvin K. Ogilvie
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 22:59
CPC Classifications
C; C07; C07H; C07H21/00; C07H1/00; C07H1/02; C07H19/00; C07H19/02; C07H19/04; C07H19/16; C07H19/20
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
160 words
A process for preparing lyophilized phosphorylated nucleotides of general formula 1 ##STR5## where R is hydrogen, oxy-lower alkyl, oxyphenyl, oxybenzyl, oxysilyl, oxy-lower alkylsilyl, oxytrityl, oxy-loweralkoxytrityl or a halophosphite group ##STR6## in which X is fluorine, chlorine, bromine or iodine and R 1 is a chemical protecting group for hydroxyl and selected from lower alkyl, phenyl, benzyl, silyl, lower alkylsilyl, trityl and lower-alkoxytrityl; R 2 and R 3 are independently selected from hydrogen, lower alkyl, phenyl, benzyl, silyl, lower alkyl silyl, trityl, lower-alkoxytrityl and halophosphite group R 1 O--P--X with the proviso that one of R, R 2 and R 3 is a halophosphite group; B' is a base selected from guanine, adenine, uracil, cytosine and thymine; which comprises forming a solution of said nucleotide in dioxane solvent, subjecting the solution to low pressure conditions to cause sublimation of the dioxane solvent, and recovering lyophilized nucleotide compound of formula 1 in light, divided form, readily soluble in pyridine at room temperatures.
Claim 2
Dependent Claim
160 words
A process for preparing lyophilized phosphorylated nucleotides of general formula 1 ##STR5## where R is hydrogen, oxy-lower alkyl, oxyphenyl, oxybenzyl, oxysilyl, oxy-lower alkylsilyl, oxytrityl, oxy-loweralkoxytrityl or a halophosphite group ##STR6## in which X is fluorine, chlorine, bromine or iodine and R 1 is a chemical protecting group for hydroxyl and selected from lower alkyl, phenyl, benzyl, silyl, lower alkylsilyl, trityl and lower-alkoxytrityl; R 2 and R 3 are independently selected from hydrogen, lower alkyl, phenyl, benzyl, silyl, lower alkyl silyl, trityl, lower-alkoxytrityl and halophosphite group R 1 O--P--X with the proviso that one of R, R 2 and R 3 is a halophosphite group; B' is a base selected from guanine, adenine, uracil, cytosine and thymine; which comprises forming a solution of said nucleotide in dioxane solvent, subjecting the solution to low pressure conditions to cause sublimation of the dioxane solvent, and recovering lyophilized nucleotide compound of formula 1 in light, divided form, readily soluble in pyridine at room temperatures.
Claim 3
Dependent Claim
158 words
A process for preparing lyophilized phosphorylated nucleotide compounds of general formula I given in claim 1, which comprises reacting, in liquid solution in dioxane, a nucleoside compound of general formula (III) ##STR7## wherein R 4 is hydrogen, hydroxyl, oxy-lower alkyl, oxyphenyl, oxybenzyl, oxysilyl, oxy-lower alkylsilyl, oxytrityl and oxy-loweralkoxytrityl; R 5 and R 6 are independently selected from hydrogen lower alkyl, phenyl, benzyl, silyl, lower alkylsilyl, trityl and lower alkoxytrityl, with the proviso that only one of R 4 , OR 5 and OR 6 represents hydroxyl, and B' has the same meanings as in formula I, with a phosphodihalidite compound of formula X 2 .PO.R 7 where X is fluorine, chlorine, bromine or iodine, and R 7 is lower alkyl, to form a compound of general formula I above, and subjecting the reaction solution so formed to cooling and low pressure conditions to sublime off the dioxane solvent, and recovering dry stable lyophilized nucleotide product of formula I.
Claim 4
Dependent Claim
18 words
The process of claim 2 wherein the phosphodihalidite compound is methyldichlorophosphite, of formula CH 3 --PO--Cl 2 .
Claim 5
Dependent Claim
30 words
The process of claim 3 wherein R 4 and R 5 represent hydrogen and R 6 is selected from lower alkyl, phenyl, benzyl, silyl, lower alkylsilyl, trityl, and lower alkoxytrityl.
Claim 6
Dependent Claim
41 words
The process of claim 3 wherein R 4 is selected from oxy-loweralkyl, oxyphenyl, oxybenzyl, oxysilyl, oxy-lower alkylsilyl, oxytrityl and oxy-loweralkoxytrityl, R 6 is selected from lower alkyl, phenyl, benzyl, silyl, lower alkylsilyl, trityl, and lower alkoxytrityl and R 5 is hydrogen.
References Cited (Backward Citations)
US3338882A
Cited
US3433782A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-09-12 | 1987-03-14 | 1987-09-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-09-12 | 1991-03-14 | 1991-09-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-09-12 | 1995-03-14 | 1995-09-13 | $7700.00 | Window Closed / Lapsed |