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US 5,700,922 AGranted
PNA-DNA-PNA chimeric macromolecules
AssigneeIsis Pharmaceuticals, Inc.
InventorPhillip Dan Cook
Life of this patent
Estimating the term…
Abstract
Macromolecules are provided that have increased nuclease resistance, increasing binding affinity to a complementary strand, and that activate RNase H enzyme. The macromolecules have the structure PNA-DNA-PNA where the DNA portion is composed of subunits of 2'-deoxy-erythro-pentofuranosyl nucleotides and the PNA portions are composed of subunits of peptide nucleic acids. Such macromolecules are useful for diagnostics and other research purposes, for modulating protein in organisms, and for the diagnosis, detection and treatment of other conditions susceptible to therapeutics.
Claims23
A macromolecule of the structure:
- PNA-DNA-PNA
- wherein:
- said DNA of said PNA-DNA-PNA comprises at least one 2'-deoxynucleotide;
- and
- each of said PNAs of said PNA-DNA-PNA comprise at least one peptide nucleic acid subunit having formula (I):
- ##STR6## wherein n is at least 2,
- each of L.sup.1 and L.sup.n is independently selected from the group consisting of hydrogen, hydroxy, (C.sub.1 -C.sub.4)alkanoyl, nucleobases, aromatic moieties, DNA intercalators, nucleobase-binding groups, heterocyclic moieties, and reporter ligands, at least one of L.sup.1 to L.sup.n being a nucleobase, a DNA intercalator, or a nucleobase-binding group;
- each of R.sup.3 and R.sup.4 is independently selected from the group consisting of hydrogen, (C.sub.1 -C.sub.4)alkyl, hydroxy- or alkoxy- or alkylthio-substituted (C.sub.1 -C.sub.4)alkyl, hydroxy, alkoxy, alkylthio and amino;
- each of C.sup.1 to C.sup.n is (CR.sup.6 R.sup.7).sub.y where R.sup.6 is hydrogen and R.sup.7 is selected from the group consisting of the side chains of naturally occurring alpha amino acids, or R.sup.6 and R.sup.7 are independently selected from the group consisting of hydrogen, (C.sub.2 -C.sub.6)alkyl, aryl, aralkyl, heteroaryl, hydroxy, (C.sub.1 -C.sub.6)alkoxy, (C.sub.1 -C.sub.6)alkylthio, NR.sup.3 R.sup.4 and SR.sup.5, where R.sup.3 and R.sup.4 are as defined above, and R.sup.5 is hydrogen, (C.sub.1 -C.sub.6)alkyl, hydroxy-, alkoxy-, or alkylthio-substituted (C.sub.1 -C.sub.6)alkyl, or R.sup.6 and R.sup.7, together, are alicyclic or heterocyclic;
- each of D.sup.1 to D.sup.n is (CR.sup.6 R.sup.7).sub.z where R.sup.6 and R.sup.7 are as defined above;
- each of y and z is zero or an integer from 1 to 10, the sum y+z being greater than 2 but not more than 10;
- each of G.sup.1 to G.sup.n-1 is --NR.sup.3 CO--, --NR.sup.3 CS--, --NR.sup.3 SO-- --NR.sup.3 SO.sub.2 --, --CONR.sup.3 --, --CSNR.sup.3 --, --SONR.sup.3 -- or --SO.sub.2 NR.sub.3 -- where R.sup.3 is as defined above;
- each pair of A.sup.1 to A.sup.n and B.sup.1 to B.sup.n are selected such that:
- (a) A is a group of formula (IIa), (IIb) or (IIc) and B is N or R.sup.3 N.sup.+ ;
- or
- (b) A is a group of formula (IId) and B is CH;
- ##STR7## where:
- X is O, S, Se, NR.sup.3, CH.sub.2 or C(CH.sub.3).sub.2 ;
- Y is a single bond, O, S or NR.sup.4 ;
- each of p and q is zero or an integer from 1 to 5, the sum p+q being not more than 10;
- each of r and s is zero or an integer from 1 to 5, the sum r+s being not more than 10;
- each R.sup.1 and R.sup.2 is independently selected from the group consisting of hydrogen, (C.sub.1 -C.sub.4)alkyl which may be hydroxy- or alkoxy- or alkylthio-substituted, hydroxy, alkoxy, alkylthio, amino and halogen;
- Q is --CO.sub.2 H, --CONR'R", --SO.sub.3 H or --SO.sub.2 NR'R" or an activated derivative of --CO.sub.2 H or --SO.sub.3 H;
- and
- I is --NR'"R"" or --NR'"C(O)R"";
- where:
- R', R", R'", and R"" are independently selected from the group consisting of hydrogen, alkyl, amino protecting groups, reporter ligands, intercalators, chelators, peptides, carbohydrates, lipids, steroids, nucleosides, nucleotides, nucleotide diphosphates, nucleotide triphosphates, oligonucleotides, oligonucleosides and soluble and non-soluble polymers, provided that at least one R' is said DNA of said PNA-DNA-PNA and at least one R"" is said DNA of said PNA-DNA-PNA.
24 more elements
A macromolecule of the structure:
- PNA-DNA or DNA-PNA
- wherein:
- said DNA of said PNA-DNA or DNA-PNA comprises at least one 2'-deoxynucleotide;
- and
- said PNA of said PNA-DNA or DNA-PNA comprises at least one peptide nucleic acid subunit having formula (I):
- ##STR8## wherein:
- n is at least 2,
- each of L.sup.1 to L.sup.n is independently selected from the group consisting of hydrogen, hydroxy, (C.sub.1 -C.sub.4)alkanoyl, nucleobases, aromatic moieties, DNA intercalators, nucleobase-binding groups, heterocyclic moieties, and reporter ligands, at least one of L.sup.1 to L.sup.n being a nucleobase, a DNA intercalator, or a nucleobase-binding group;
- each of R.sup.3 and R.sup.4 is independently selected from the group consisting of hydrogen, (C.sub.1 -C.sub.4)alkyl, hydroxy- or alkoxy- or alkylthio-substituted (C.sub.1 -C.sub.4)alkyl, hydroxy, alkoxy, alkylthio and amino;
- each of C.sup.1 to C.sup.n is (CR.sup.6 R.sup.7).sub.y where R.sup.6 is hydrogen and R.sup.7 is selected from the group consisting of the side chains of naturally occurring alpha amino acids, or R.sup.6 and R.sup.7 are independently selected from the group consisting of hydrogen, (C.sub.2 -C.sub.6)alkyl, aryl, aralkyl, heteroaryl, hydroxy, (C.sub.1 -C.sub.6)alkoxy, (C.sub.1 -C.sub.6)alkylthio, NR.sup.3 R.sup.4 and SR.sup.5, where R.sup.3 and R.sup.4 are as defined above, and R.sup.5 is hydrogen, (C.sub.1 -C.sub.6)alkyl, hydroxy-, alkoxy-, or alkylthio-substituted (C.sub.1 -C.sub.6)alkyl, or R.sup.6 and R.sup.7, together, are alicyclic or heterocyclic;
- each of D.sup.1 to D.sup.n is (CR.sup.6 R.sup.7).sub.z where R.sup.6 and R.sup.7 are as defined above;
- each of y and z is zero or an integer from 1 to 10, the sum y+z being greater than 2 but not more than 10;
- each of G.sup.1 to G.sup.n-1 is --NR.sup.3 CO--, --NR.sup.3 CS--, --NR.sup.3 SO-- --NR.sup.3 SO.sub.2 --, --CONR.sup.3 --, --CSNR.sup.3 --, --SONR.sup.3 -- where R.sup.3 is as defined above;
- each pair of A.sup.1 to A.sup.n and B.sup.1 to B.sup.n are selected such that:
- (a) A is a group of formula (IIa), (IIb) or (IIc) and B is N or R.sup.3 N.sup.+ ;
- or
- (b) A is a group of formula (IId) and B is CH;
- ##STR9## where X is O, S, Se, NR.sup.3, CH.sub.2 or C(CH.sub.3).sub.2 ;
- Y is a single bond, O, S or NR.sup.4 ;
- each of p and q is zero or an integer from 1 to 5, the sum p+q being not more than 10;
- each of r and s is zero or an integer from 1 to 5, the sum r+s being not more than 10;
- each R.sup.1 and R.sup.2 is independently selected from the group consisting of hydrogen, (C.sub.1 -C.sub.4)alkyl which may be hydroxy- or alkoxy- or alkylthio-substituted, hydroxy, alkoxy, alkylthio, amino and halogen;
- Q is --CO.sub.2 H, --CONR'R", --SO.sub.3 H or --SO.sub.2 NR'R" or an activated derivative of --CO.sub.2 H or --SO.sub.3 H;
- and
- I is --NR'"R"" or --NR'"C(O)R"";
- where:
- R', R", R'", and R"" are independently selected from the group consisting of hydrogen, alkyl, amino protecting groups, reporter ligands, intercalators, chelators, peptides, carbohydrates, lipids, steroids, nucleosides, nucleotides, nucleotide diphosphates, nucleotide triphosphates, oligonucleotides, oligonucleosides and soluble and non-soluble polymers, provided that R' is said DNA of said PNA-DNA or DNA-PNA or R"" is said DNA of said PNA-DNA or DNA-PNA.
24 more elements
A macromolecule of the structure:
- PNA-(amide link)-DNA-(amide link)-PNA:
- wherein:
- said DNA of said structure comprises at least one 2'-deoxynucleotide;
- each of said PNAs of said structure comprise at least one peptide nucleic acid subunit;
- and
- each of said amide links of said structure includes an amide linkage of the structure:
- --C(.dbd.O)--NH-- or --NH--C(.dbd.O)--.
4 more elements
Dependencies are inferred from the claim text.