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US5736626A
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Solid support reagents for the direct synthesis of 3'-labeled polynucleotides
Assignee: Applied Biosystems LLC
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Filing: 1996-01-29
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Granted: 1998-04-07
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Expires: 2016-01-29
Abstract
Abstract The compounds of the invention are exemplified by the class of diglycolate synthesis supports particularly useful as support reagents for the direct synthesis of 3'-labeled polynucleotides. Generally, the compounds of the invention have the structure protecting group, e.g., 4,4'-dimethoxytrityl; L1 is a linker for connecting a 3'-terminal nitrogen to carbon; L2 and L3 are linkers for connecting oxygen and carbon; W is a solid support, e.g., CPG or polystyrene; L4 is a linker for connecting the solid support to nitrogen; R1 and R2 are nitrogen substituents, e.g., hydrogen, lower alkyl, nitrogen protecting group, or label; and R3 through R7 are carbon substituents, e.g., hydrogen or lower alkyl. In a first particularly preferred embodiment, the synthesis supports of the invention are exemplified by compounds having the structure <IMAGE> where DMT is 4,4'-dimethoxytrityl and W is polystyrene. In a second particularly preferred embodiment, the synthesis supports of the invention are exemplified by compounds having the structure <IMAGE> where DMT and W are defined above.
Inventors
- 👤 Khairuzzaman Bashar Mullah
- 👤 William A. Andrus
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 20:02
CPC Classifications
C; C07; C07H; C07H21/00; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55; Y10; Y10S; Y10S435/00; Y10S435/81
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
108 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR14## where T is an acid-cleavable hydroxyl protecting group; L 1 is a linker for connecting a 3'-terminal nitrogen to carbon; L 2 and L 3 are linkers for connecting oxygen and carbon; W is a solid support; L 4 is a linker for connecting the solid support to nitrogen; R 1 and R 2 are nitrogen substituents individually selected from the group consisting of hydrogen, lower alkyl, nitrogen protecting group, or label; and R 3 through R 7 are carbon substituents individually selected from the group consisting of hydrogen or lower alkyl.
Claim 2
Dependent Claim
108 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR14## where T is an acid-cleavable hydroxyl protecting group; L 1 is a linker for connecting a 3'-terminal nitrogen to carbon; L 2 and L 3 are linkers for connecting oxygen and carbon; W is a solid support; L 4 is a linker for connecting the solid support to nitrogen; R 1 and R 2 are nitrogen substituents individually selected from the group consisting of hydrogen, lower alkyl, nitrogen protecting group, or label; and R 3 through R 7 are carbon substituents individually selected from the group consisting of hydrogen or lower alkyl.
Claim 3
Dependent Claim
9 words
The support of claim 1 wherein W is CPG.
Claim 4
Dependent Claim
12 words
The support of claim 2 wherein L 4 has the structure ##STR15##
Claim 5
Dependent Claim
10 words
The support of claim 1 wherein W is nonswellable polystyrene.
Claim 6
Dependent Claim
10 words
The support of claim 4 wherein L 4 is methylene.
Claim 7
Dependent Claim
9 words
The support of claim 1 wherein T is 4,4'-dimethoxytrityl.
Claim 8
Dependent Claim
19 words
The support of claim 1 wherein L 1 has the structure ##STR16## where n ranges from 0 to 10.
Claim 9
Dependent Claim
12 words
The support of claim 7 wherein n ranges from 4 to 6.
Claim 10
Dependent Claim
14 words
The support of claim 1 wherein L 2 and L 3 are each methylene.
Claim 11
Dependent Claim
11 words
The support of claim 1 wherein R 2 is tetramethyl rhodamine.
Claim 12
Dependent Claim
12 words
The support of claim 1 wherein L 3 includes a disulfide bond.
Claim 13
Dependent Claim
25 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR17## where DMT is 4,4'-dimethoxytrityl and W is non-swellable polystyrene.
Claim 14
Dependent Claim
25 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR17## where DMT is 4,4'-dimethoxytrityl and W is non-swellable polystyrene.
Claim 15
Dependent Claim
24 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR18## where DMT is 4,4'-dimethoxytrityl and W is CPG.
Claim 16
Dependent Claim
24 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR18## where DMT is 4,4'-dimethoxytrityl and W is CPG.
Claim 17
Dependent Claim
25 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR19## where DMT is 4,4'-dimethoxytrityl and W is non-swellable polystyrene.
Claim 18
Dependent Claim
25 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR19## where DMT is 4,4'-dimethoxytrityl and W is non-swellable polystyrene.
Claim 19
Dependent Claim
24 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR20## where DMT is 4,4'-dimethoxytrityl and W is CPG.
Claim 20
Dependent Claim
24 words
A synthesis support for the synthesis of 3'-labeled polynucleotides comprising a compound of the formula: ##STR20## where DMT is 4,4'-dimethoxytrityl and W is CPG.
Claim 21
Independent Claim
178 words
A method for synthesizing 3'-labeled polynucleotides comprising the steps of: (a) providing a synthesis support comprising a compound of the formula ##STR21## where, T is an acid-cleavable hydroxyl protecting group; L 1 is a linker for connecting a 3'-terminal nitrogen to carbon; L 2 and L 3 are linkers for connecting oxygen and carbon; W is a solid support; L 4 is a linker for connecting the solid support to nitrogen; R 1 and R 2 are nitrogen substituents individually selected from the group consisting of hydrogen, lower alkyl, nitrogen protecting group, or label; and R 3 through R 7 are carbon substituents individually selected from the group consisting of hydrogen or lower alkyl; (b) treating the solid support with acid to remove the add-cleavable hydroxyl protecting group; (c) adding a protected nucleoside monomer and a weak acid, forming a linkage; (d) capping unreacted sites on the solid support; (e) adding an oxidizing reagent; (f) repeating steps (b)-(e) until the polynucleotide chain elongation is complete. (g) cleaving the polynucleotide from the solid support; and (h) deprotecting the polynueleotide.
Claim 22
Independent Claim
178 words
A method for synthesizing 3'-labeled polynucleotides comprising the steps of: (a) providing a synthesis support comprising a compound of the formula ##STR21## where, T is an acid-cleavable hydroxyl protecting group; L 1 is a linker for connecting a 3'-terminal nitrogen to carbon; L 2 and L 3 are linkers for connecting oxygen and carbon; W is a solid support; L 4 is a linker for connecting the solid support to nitrogen; R 1 and R 2 are nitrogen substituents individually selected from the group consisting of hydrogen, lower alkyl, nitrogen protecting group, or label; and R 3 through R 7 are carbon substituents individually selected from the group consisting of hydrogen or lower alkyl; (b) treating the solid support with acid to remove the add-cleavable hydroxyl protecting group; (c) adding a protected nucleoside monomer and a weak acid, forming a linkage; (d) capping unreacted sites on the solid support; (e) adding an oxidizing reagent; (f) repeating steps (b)-(e) until the polynucleotide chain elongation is complete. (g) cleaving the polynucleotide from the solid support; and (h) deprotecting the polynueleotide.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-04-06 | 2001-10-06 | 2002-04-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-04-06 | 2005-10-06 | 2006-04-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-04-06 | 2009-10-06 | 2010-04-07 | $7700.00 | Window Closed / Lapsed |