Antisense antibacterial cell division composition and method

• US 7,049,431 B2
• Filed: 01/04/2001
• Issued: 05/23/2006
• Est. Priority Date: 01/04/2000
• Status: Expired due to Term

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Avi Biopharma, Inc.
• Inventors: Iversen, Patrick L.
• Primary Examiner(s): McGarry, Sean
• Assistant Examiner(s): ZARA, JANE J
• Application Number:
  US09/754,468
• Publication Number:
  US 20050192237A1
• Time in Patent Office: 1,965 Days
• Field of Search: 435/6, 435/91.1, 435/455, 435/458, 435/91.31, 514/44, 536/23.1, 536/24.5, 536/24.31
• US Class Current: 536/24.5
• CPC Class Codes: A23K 20/195 Antibiotics
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring

Abstract

Antisense oligomers directed to bacterial cell division and cell cycle-encoding nucleic acids are capable of selectively modulating the biological activity thereof, and are useful in treatment and prevention of bacterial infection. The antisense oligomers are substantially uncharged, and contain from 8 to 40 nucleotide subunits, including a targeting nucleic acid sequence at least 10 nucleotides in length which is effective to hybridize to (i) a bacterial tRNA or (ii) a target sequence, containing a translational start codon, within a bacterial nucleic acid which encodes a protein associated with cell division or the cell cycle. Such proteins include zipA, sulA, secA, dicA, dicB, dicC, dicF, ftsA, ftsI, ftsN, ftsK, ftsL, ftsQ, ftsW, ftsZ, murC, murD, murE, murF, murG, minC, minD, minE, mraY, mraW, mraZ, seqA, ddlB, carbamate kinase, D-ala D-ala ligase, topoisomerase, alkyl hydroperoxide reductase, thioredoxin reductase, dihydrofolate reductase, and cell wall enzyme.

Claims

1. A substantially uncharged antisense oligomer containing 10 to 40 morpholino subunits, each of said subunits supporting a base-pairing moiety effective to bind by Watson-Crick base pairing to a respective nucleotide base,

wherein said base-pairing moieties include a targeting nucleic acid sequence having the sequence presented as SEQ ID NO:

47,

and wherein adjacent subunits are joined by uncharged linkages selected from the group consisting of uncharged phosphoramidate and phosphorodiamidate, or by charged linkages selected from the group consisting of charged phosphoramidate and phosphorodiamidate, the ratio of uncharged linkages to charged linkages in the oligomer being at least 4;

1.

2. The oligomer of claim 1, wherein each said uncharged linkage is a phosphorodiamidate linkage as represented by —

P(═

O)(NR₂)—

O—

, where R is hydrogen or methyl.

3. The oligomer of claim 2, wherein each said linkage in said oligomer is an uncharged phosphorodiamidate linkage as represented by —

P(═

O)(NR₂)—

O—

, where R is hydrogen or methyl.

4. The oligomer of claim 1, wherein the length of said oligomer is less than 30 subunits.

5. The oligomer of claim 1, wherein the length of said oligomer is less than 25 subunits.

6. The oligomer of claim 1, wherein the length of said oligomer is 20 subunits.