Antisense antibacterial compounds and methods
AssigneesBoard of Regents, The University of Texas System, Oregon State University
InventorsBruce L. Geller, David Greenberg
Life of this patent
Estimating the term…
Abstract
Provided are antisense oligomers targeted against or genes associated with a biochemical pathway and/or cellular process, and related compositions and methods of using the oligomers and compositions to treat an infected mammalian subject, for example, as primary antimicrobials or as adjunctive therapies with classic antimicrobials.
Claims46
An antisense morpholino oligomer, composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′-exocyclic carbon of an adjacent subunit, and having (a) about 10-40 nucleotide bases, and (b) a targeting sequence of sufficient length and complementarity to specifically hybridize to a bacterial mRNA target sequence that encodes a protein associated with a biochemical pathway and/or cellular process, or a protein associated with antibiotic resistance, where the oligomer is conjugated to a cell-penetrating peptide (CPP).
The antisense morpholino oligomer of formula (I):
- or a pharmaceutically acceptable salt thereof,
- where each Nu is a nucleobase which taken together forms a targeting sequence;
- X is an integer from 9 to 38;
- T is selected from OH and a moiety of the formula:
- where each R4 is independently C1-C6 alkyl, and R5 is selected from an electron pair and H, and R6 is selected from OH, —N(R7)CH2C(O)NH2, and a moiety of the formula:
- where:
- R7 is selected from H and C1-C6 alkyl, and
- R8 is selected from G, —C(O)—R9OH, acyl, trityl, and 4-methoxytrityl, where:
- R9 is of the formula —(O-alkyl)y- where y is an integer from 3 to 10 and each of
- the y alkyl groups is independently selected from C2-C6 alkyl;
- each instance of R1 is —N(R10)2R11 where each R10 is independently C1-C6 alkyl, and R11 is
- selected from an electron pair and H;
- R2 is selected from H, G, acyl, trityl, 4-methoxytrityl, benzoyl, stearoyl, and a moiety of the formula:
- where L is selected from —C(O)(CH2)6C(O)— and —C(O)(CH2)2S2(CH2)2C(O)—, and each R12 is of the formula —(CH2)2OC(O)N(R14)2 where each R14 is of the formula —(CH2)6NHC(═NH)NH2;
- and
- R3 is selected from an electron pair, H, and C1-C6 alkyl,
- where G is a cell penetrating peptide (“CPP”) and linker moiety selected from —C(O)(CH2)5NH—CPP, —C(O)(CH2)2NH—CPP, —C(O)(CH2)2NHC(O)(CH2)5NH—CPP, and —C(O)CH2NH—CPP, or G is of the formula:
- where the CPP is attached to the linker moiety by an amide bond at the CPP carboxy terminus, with the proviso that only one instance of G is present,
- where the targeting sequence specifically hybridizes to a bacterial mRNA target sequence that encodes a protein selected from at least one of a protein associated with a biochemical pathway and/or cellular process and antibiotic resistance.
A pharmaceutical composition, comprising a pharmaceutically acceptable carrier and an antisense morpholino oligomer, wherein the antisense morpholino oligomer is composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′-exocyclic carbon of an adjacent subunit, and having (a) about 10-40 nucleotide bases, and (b) a targeting sequence of sufficient length and complementarity to specifically hybridize to a bacterial mRNA target sequence that encodes a protein associated with a biochemical pathway and/or cellular process, or a protein associated with antibiotic resistance, where the oligomer is conjugated to a cell-penetrating peptide (CPP).
A method of reducing expression and activity of a protein selected from at least at least one of a protein associated with a biochemical pathway and/or cellular process and antibiotic resistance in a bacterium, comprising contacting the bacterium with an antisense morpholino oligomer, wherein the antisense morpholino oligomer is composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′-exocyclic carbon of an adjacent subunit, and having (a) about 10-40 nucleotide bases, and (b) a targeting sequence of sufficient length and complementarity to specifically hybridize to a bacterial mRNA target sequence that encodes a protein associated with a biochemical pathway and/or cellular process, or a protein associated with antibiotic resistance, where the oligomer is conjugated to a cell-penetrating peptide (CPP).
The method of 44, where the antimicrobial agent is selected from one or more of a β-lactam antibiotic, an aminoglycoside antibiotic, and a polymyxin.
Dependencies are inferred from the claim text.