Patent US830513A successfully refreshed from Google Patents.
Patent US5296350A successfully refreshed from Google Patents.
US5296350A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Ion triggered alkylation of biological targets by silyloxy aromatic agents
Assignee: Research Foundation of the State University of New York
•
Filing: 1990-10-31
•
Granted: 1994-03-22
•
Expires: 2010-10-31
Abstract
Abstract A sequence directed reagent is constructed by conjugating a methyl silyloxy aromatic derivative to a hexamethylamino linker attached to either the 5' or 3' terminus of an oligonucleotide. Annealing this modified fragment of DNA to its complementary sequence allows for target modification subsequent to ionic activation. The product of this reaction is a covalent crosslink between the reagent and target strands resulting from an alkylation of DNA by the activated silyloxy aromatic derivative. In a preferred embodiment, a nitrophenyl group is attached to the methyl group of the silyloxy aromatic derivative. This reagent is similarly linked to an oligonucleotide probe. Activation of this probe linked alkylating agent by an ionic signal, (X) which may naturally occur, or may be introduced into the media containing the target molecule, such as by the introduction of a salt (MX).
Inventors
- 👤 Steven E. Rokita
- 👤 Tianhu Li
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 11:58
CPC Classifications
C; C07; C07H; C07H21/00; C07F; C07F7/00; C07F7/02; C07F7/08; C07F7/18; C07F7/1804; C07H23/00
Claims Analysis
Total Claims: 33
Claim 1
Independent Claim
178 words
A silyloxy aromatic probe having the formula: ##STR18## wherein when R 1 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 2 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 3 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 4 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 5 =--Osi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; wherein said R 6 , R 7 , R 8 =various alkyl or aromatic groups; X leaving group; wherein said R 9 , is H, or an organic compound, such as an aliphatic or alkyl group L is a linking group for attachment to a probe which may be positioned at any carbon atom of the ring; and Pr is a probe for binding to a target molecule.
Claim 2
Dependent Claim
178 words
A silyloxy aromatic probe having the formula: ##STR18## wherein when R 1 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 2 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 3 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 4 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 5 =--Osi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; wherein said R 6 , R 7 , R 8 =various alkyl or aromatic groups; X leaving group; wherein said R 9 , is H, or an organic compound, such as an aliphatic or alkyl group L is a linking group for attachment to a probe which may be positioned at any carbon atom of the ring; and Pr is a probe for binding to a target molecule.
Claim 3
Dependent Claim
61 words
A silyloxy aromatic probe according to claim 1, wherein said silyloxyl aromatic probe alkylates a target molecule after ionic activation; and wherein X is selected from the group consisting of Cl, F, I, OCOR, OH, OSO 2 CH 3 , OSO 2 C 6 H 4 CH 3 -p, OR, OCONHR, OCONHCH 2 CH 2 R, Br, phenol, nitro-phenol and thio-phenol.
Claim 4
Dependent Claim
96 words
A silyloxy aromatic probe according to claim 2, wherein L comprises a chain --R 10 --R 11 --R 12 --, in which R 10 is selected from the group consisting of NH, S, O and CH 2 , in which R 11 comprises a stable spacer group between R 11 and R 12 , and in which R 12 is selected from the group consisting of NH 2 , SH, OH and COOH; and wherein Pr is a probe that includes a localizing moiety, such as an oligonucleotide that preferentially localizes to an DNA or RNA.
Claim 5
Dependent Claim
21 words
A silyloxy aromatic probe according to claim 3 wherein R 6 , R 7 and R 8 comprises a t-butyl moiety.
Claim 6
Dependent Claim
13 words
A silyloxy aromatic probe according to claim 3, wherein Pr is an oligonucleotide.
Claim 7
Dependent Claim
14 words
A silyloxy aromatic probe according to claim 5, wherein Pr is a DNA strand.
Claim 8
Dependent Claim
19 words
A silyloxy aromatic probe according to claim 5, wherein said oligonucleotide is linked to L by its 5' terminus.
Claim 9
Dependent Claim
19 words
A silyloxy aromatic probe according to claim 5, wherein the oligonucleotide is linked to L by its 3' terminus.
Claim 10
Dependent Claim
26 words
A silyloxy aromatic probe according to claim 5, further comprising a modified base on said oligonucleotide suitable for linking to R 12 at said modified base.
Claim 11
Dependent Claim
28 words
A silyloxy aromatic probe according to claim 5, further comprising a modified phosphoribose backbone on said oligonucleotide suitable for linking to R 12 at said modified phosphodeoxyribose backbone.
Claim 12
Dependent Claim
71 words
A silyloxy aromatic probe composition having the formula: ##STR19## wherein R 2 =--OSi(R 6 R 7 R 8 ), R 3 =--CH 2 X; said R 6 , R 7 , R 8 =various alkyl or aromatic groups; X=leaving group; L is a linking group for to a probe which may be positioned at any carbon atom of the ring; and P is a probe for binding to a target molecule.
Claim 13
Dependent Claim
71 words
A silyloxy aromatic probe composition having the formula: ##STR19## wherein R 2 =--OSi(R 6 R 7 R 8 ), R 3 =--CH 2 X; said R 6 , R 7 , R 8 =various alkyl or aromatic groups; X=leaving group; L is a linking group for to a probe which may be positioned at any carbon atom of the ring; and P is a probe for binding to a target molecule.
Claim 14
Dependent Claim
91 words
A silyloxy aromatic probe according to claim 11, wherein L comprises a chain --R 10 --R 11 --R 12 --, in which R 10 is selected from the group consisting of N, S, O and CH 2 , in which R 11 comprises a stable spacer group between R 10 and R 11 , and in which R 12 is selected from the group consisting of NH 2 , SH, OH and COOH; and wherein Pr is a probe that includes a localizing moiety, such as an oligonucleotide DNA or RNA.
Claim 15
Dependent Claim
21 words
A silyloxy aromatic probe according to claim 12, wherein R 6 , R 7 and R 8 comprises a t-butyl moiety.
Claim 16
Dependent Claim
13 words
A silyloxy aromatic probe according to claim 12, wherein Pr is an oligonucleotide.
Claim 17
Dependent Claim
14 words
A silyloxy aromatic probe according to claim 12, wherein Pr is a DNA strand.
Claim 18
Dependent Claim
19 words
A silyloxy aromatic probe according to claim 14, wherein said oligonucleotide is linked to L by its 5' terminus.
Claim 19
Dependent Claim
19 words
A silyloxy aromatic probe according to claim 14, wherein said oligonucleotide is linked to L by its 3' terminus.
Claim 20
Dependent Claim
26 words
A silyloxy aromatic probe according to claim 14, further comprising a modified base on said oligonucleotide suitable for linking to R 12 at said modified base.
Claim 21
Dependent Claim
28 words
A silyloxy aromatic probe according to claim 14, further comprising a modified phosphoribose backbone on said oligonucleotide suitable for linking to R 12 at said modified phosphoribose backbone.
Claim 22
Dependent Claim
18 words
A silyloxy aromatic probe recited in claim 13, wherein X comprises C 6 H 4 NO 3 .
Claim 23
Dependent Claim
110 words
A process for selectively alkylating a target molecule, comprising steps of: a) providing a probe for recognizing a predetermined binding site on a target molecule; b) providing a silyloxy aromatic derivative for linking to the probe; c) linking the probe to the silyloxy aromatic derivative to form a targeted alkylating agent; d) introducing the targeted alkylating agent to a system containing a target molecule, whereby the probe associates with the target molecule, localizing the linked silyloxy aromatic derivative near the target molecule; and e) activating the targeted alkylating agent, thereby causing covalent binding between the linked aromatic derivative proximal to the association site of the probe with the target molecule.
Claim 24
Dependent Claim
110 words
A process for selectively alkylating a target molecule, comprising steps of: a) providing a probe for recognizing a predetermined binding site on a target molecule; b) providing a silyloxy aromatic derivative for linking to the probe; c) linking the probe to the silyloxy aromatic derivative to form a targeted alkylating agent; d) introducing the targeted alkylating agent to a system containing a target molecule, whereby the probe associates with the target molecule, localizing the linked silyloxy aromatic derivative near the target molecule; and e) activating the targeted alkylating agent, thereby causing covalent binding between the linked aromatic derivative proximal to the association site of the probe with the target molecule.
Claim 25
Dependent Claim
183 words
A process as recited in claim 21, wherein the targeted alkylating agent has the molecular formula: ##STR20## wherein when R 1 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 2 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 3 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; when R 4 =--OSi(R 6 R 7 R 8 ), then R 1 , R 3 and/or R 5 is=--CHR 9 X; when R 5 =--OSi(R 6 R 7 R 8 ), then R 2 and/or R 4 is=--CHR 9 X; wherein said R 6 , R 7 , R 8 =various alkyl or aromatic groups; said R 9 is H, or an organic group, such as an aliphatic oralkyl group X=leaving group; L is a linking group for attachment to a probe which may be positioned at any carbon atom of the ring; and Pr is a probe for binding to a target molecule.
Claim 26
Dependent Claim
39 words
A process as recited in claim 22, wherein said linking step (c) further comprises adapting the silyloxy aromatic derivative by the addition of an acidic linking group suitably modified for linking the silyloxy aromatic derivative to the probe molecule.
Claim 27
Dependent Claim
35 words
A process as recited in claim 23, wherein said probe is an oligonucleotide, and further comprising the step of: suitably modifying a base of said oligonucleotide probe for linking to L at the modified base.
Claim 28
Dependent Claim
37 words
A process as recited in claim 23, wherein said probe is an oligonucleotide, and further comprising the step of: suitably modifying a phosphoribose backbone of said oligonucleotide probe for linking to L at the modified phosphoribose backbone.
Claim 29
Dependent Claim
18 words
A process as recited in claim 23, wherein said activating step comprises introduction of an ionic activating signal.
Claim 30
Dependent Claim
27 words
A process as recited in claim 23, wherein said adapting step further includes brominating the X group, and said activating step comprises activation with an ionic signal.
Claim 31
Dependent Claim
36 words
A process recited in claim 27, wherein said brominating step is followed by substituting the bromine by molecules selected from the group consisting of Cl, F, OCONHCH 2 CH 2 R, Br, phenol, nitro-phenol and thio-phenol.
Claim 32
Dependent Claim
91 words
A process as recited in claim 28, wherein L comprises a chain --R 10 --R 11 --R 12 --, in which R 10 is selected from the group consisting of N, S, O and CH 2 , in which R 11 comprises a stable spacer group between R 10 and R 12 , and in which R 12 is selected from the group consisting of NH 2 , SH, OH and COOH; and wherein Pr is a probe that includes a localizing moiety, such as an oligonucleotide that preferentially localizes to.
Claim 33
Dependent Claim
15 words
A process as recited in claim 28, wherein said process is carried out in vitro.
References Cited (Backward Citations)
WO1985002628A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-03-21 | 1997-09-20 | 1998-03-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-03-21 | 2001-09-20 | 2002-03-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-03-21 | 2005-09-20 | 2006-03-22 | $7700.00 | Window Closed / Lapsed |