Human RNase H1 mutants

• US 6,855,530 B2
• Filed: 11/14/2001
• Issued: 02/15/2005
• Est. Priority Date: 11/15/2000
• Status: Expired due to Fees

• Current Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Original Assignee: ISIS Pharmaceuticals Incorporated (Ionis Pharmaceuticals, Inc.)
• Inventors: Crooke, Stanley T., Lima, Walter F., Wu, Hongjiang
• Primary Examiner(s): Patterson, Jr., Charles L.
• Application Number:
  US09/992,738
• Publication Number:
  US 20020160486A1
• Time in Patent Office: 1,189 Days
• Field of Search: 435/199
• US Class Current: 435/199
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

A human RNase H polypeptide and methods of use are provided.

Claims

1. A human RNase H1 polypeptide which comprises a mutation compared to wild type human RNase H1, wherein said mutation produces a mutant version of human RNase H1 polypeptide that consists of a deletion mutation of region I, a deletion mutation of region II, a deletion mutation of region I and region II, or a lysine-to-alanine substitution at amino acids 226 and 227 of SEQ ID NO:

3, and wherein said mutant version retains detectable cleavage activity for the RNA strand of an RNA/DNA duplex.

2. A composition comprising the human RNase H1 polypeptide of claim 1 and a pharmaceutically acceptable carrier.

3. A human H1 polypeptide which consists solely of the basic substrate binding domain which consists of region III and which retains detectable cleavage activity for the RNA strand of an RNA/DNA duplex.

4. The RNase H1 polypeptide of claim 3 which has an initial cleavage rate for the RNA strand of an RNA/DNA duplex which is at least 50% of the initial cleavage rate for a wild type human RNase H1.

5. A composition comprising the human RNase H1 polypeptide of claim 3 and a pharmaceutically acceptable carrier.