US 5,296,350 A · Ion triggered alkylation of biological targets by silyloxy aromatic agents

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).

Claims30

Dependent claims

A silyloxy aromatic probe having the formula:

##STR18## wherein when R.sub.1 =--OSi(R.sub.6 R.sub.7 R.sub.8), then R.sub.2 and/or R.sub.4 is=--CHR.sub.9 X;
when R.sub.2 =--OSi(R.sub.6 R.sub.7 R.sub.8), then R.sub.1, R.sub.3 and/or R.sub.5 is=--CHR.sub.9 X;
when R.sub.3 =--OSi(R.sub.6 R.sub.7 R.sub.8), then R.sub.2 and/or R.sub.4 is=--CHR.sub.9 X;
when R.sub.4 =--OSi(R.sub.6 R.sub.7 R.sub.8), then R.sub.1, R.sub.3 and/or R.sub.5 is=--CHR.sub.9 X;
when R.sup.5 =--Osi(R.sub.6 R.sub.7 R.sub.8), then R.sub.2 and/or R.sub.4 is=--CHR.sub.9 X;
wherein said R.sub.6, R.sub.7, R.sub.8 =various alkyl or aromatic groups;
X leaving group;
wherein said R.sub.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.

6 more elements

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.

3 more elements

Dependencies are inferred from the claim text.

Ion triggered alkylation of biological targets by silyloxy aromatic agents

Life of this patent

Ion triggered alkylation of biological targets by silyloxy aromatic agents

Life of this patent

Abstract

Claims30