Immunosuppression compound and treatment method

• US 8,501,704 B2
• Filed: 11/07/2008
• Issued: 08/06/2013
• Est. Priority Date: 11/08/2005
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Mourich, Dan V., Iversen, Patrick L., Weller, Dwight D.
• Primary Examiner(s): GIBBS, TERRA C
• Application Number:
  US12/267,437
• Publication Number:
  US 20090110689A1
• Time in Patent Office: 1,733 Days
• Field of Search: 536/23.1, 536/24.3, 536/24.33, 536/24.5, 514/44
• US Class Current: 514/44.A
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/314 Phosphoramidates
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3513 Protein; Peptide

Abstract

Provided are methods and antisense oligonucleotide analogs for suppressing an immune response in a mammalian subject, for the treatment or prevention of an autoimmune condition or transplantation rejection. The oligonucleotide analogs provided herein comprise a targeting sequence complementary to a preprocessed CTLA-4 mRNA region that spans the splice junction between intron 1 and exon 2 of the preprocessed CTLA-4 mRNA. Also provided are methods of use, in which the oligonucleotides are effective, when administered to a subject, to form within host cells, a heteroduplex structure (i) composed of the preprocessed CTLA-4 mRNA and the oligonucleotide compound, (ii) characterized by a Tm of dissociation of at least 45° C., and (iii) resulting in an increased ratio of processed mRNA encoding ligand-independent CTLA-4 to processed mRNA encoding full-length CTLA-4.

Claims

1. A composition for suppressing an immune response in a subject, comprising

(a) a pharmaceutically effective amount of a morpholino antisense oligonucleotide having between 12 and 40 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 targeting sequence that contains at least 12 contiguous bases that are complementary to SEQ ID NO;

22, the region of SEQ ID NO;

1 targeted by the overlapping sequences identified by SEQ ID NOS;

4-6, where the oligonucleotide is capable of forming with pre-processed cytotoxic T-lymphocyte antigen-4 (CTLA-4) mRNA, a heteroduplex structure characterized by a Tm of dissociation of at least 45°

C.; and

(b) a pharmaceutically acceptable carrier.

2. The composition of claim 1, wherein said morpholino subunits in the antisense oligonucleotide are joined by phosphorodiamidate linkages, in accordance with the structure:

3. The composition of claim 1, in which at least 2 and no more than half of the total number of intersubunit linkages in the antisense oligonucleotide are cationic linkages, and wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

4. The composition of claim 1, wherein the oligonucleotide is covalently linked to an arginine-rich peptide that comprises a sequence selected from SEQ ID NOS:

15-21.

5. The composition of claim 4, wherein the arginine-rich peptide comprises SEQ ID NO:

16.

6. The composition of claim 4, wherein the arginine-rich peptide comprises SEQ ID NO:

21.

7. The composition of claim 1, wherein the oligonucleotide comprises one of the overlapping sequences identified as SEQ ID NOS:

4-6.

8. The composition of claim 6, wherein the oligonucleotide comprises SEQ ID NO:

4.

9. A method of inducing immunological tolerance in a subject, comprising administering to the subject, a pharmaceutically effective amount of the compound of claim 1.

10. The method of claim 9, wherein said administering is by a parenteral route, in an amount between 5-250 mg compound/dose.

11. The method of claim 10, wherein said administering is by intravenous or intraperitoneal administration.

12. The method of claim 9, wherein the oligonucleotide has one of the overlapping sequences identified as SEQ ID NOS:

4-6.

13. The method of claim 9, wherein the oligonucleotide is covalently linked to an arginine-rich peptide that comprises a sequence selected from SEQ ID NOS:

15-21.

14. The method of claim 9, for use in delaying the onset of Type-1 diabetes in a subject.

15. The method of claim 14, wherein said administering is by a parenteral route, in an amount between 5-250 mg compound/dose.

16. The method of claim 15, wherein said administering is by intravenous or intraperitoneal administration.