Linkers designed to intercalate the double helix greatly facilitate DNA alkylation by triplex-forming oligonucleotides carrying a cyclopropapyrroloindole reactive moiety.
Triplex-forming oligonucleotides (TFOs) bind sequence-specifically in the major groove of double-stranded DNA. Cyclopropapyrroloindole (CPI), the electrophilic moiety that comprises the reactive subunit of the antibiotic CC-1065, gives hybridization-triggered alkylation at the N-3 position of adenines when bound in the minor groove of double-stranded DNA. In order to attain TFO-directed targeting of CPI, we designed and tested linkers to 'thread' DNA from the major groove-bound TFO to the minor groove binding site of CPI. Placement of an aromatic ring in the linker significantly enhanced the site-directed reaction, possibly due to a 'threading' mechanism where the aromatic ring is intercalated. All of the linkers containing aromatic rings provided efficient alkylation of the duplex target. The linker containing an acridine ring system, the strongest intercalator in the series, gave a small but clearly detectable amount of non-TFO-specific alkylation. An equivalent-length linker without an aromatic ring was very inefficient in DNA target alkylation.
Dempcy
R O
RO
Epoch Pharmaceuticals, Inc., 12277 134th Court N.E., #110, Redmond, WA 98052, USA. rdempcy@epochpharm.com
Kutyavin
I V
IV
Mills
A G
AG
Lukhtanov
E A
EA
Meyer
R B
RB
D000166:Acridines / Q000737:chemistry / Q000378:metabolism; D000225:Adenine / Q000737:chemistry / Q000378:metabolism; D000478:Alkylation; D001483:Base Sequence; D001665:Binding Sites; D003521:Cyclopropanes / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015195:Drug Design; D000080890:Duocarmycins; D007211:Indoles / Q000378:metabolism*; D007364:Intercalating Agents / Q000378:metabolism*; D007933:Leucomycins / Q000737:chemistry; D008956:Models, Chemical; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
D000166:Acridines; D003521:Cyclopropanes; D000080890:Duocarmycins; D007211:Indoles; D007364:Intercalating Agents; D007933:Leucomycins; D009838:Oligodeoxyribonucleotides; C110747:cyclopropapyrroloindole; C051802:triplex DNA; C018782:CC 1065; D004247:DNA; D000225:Adenine