IMMUNOSUPPRESSION COMPOUND AND TREATMENT METHOD

• US 20090110689A1
• Filed: 11/07/2008
• Published: 04/30/2009
• Est. Priority Date: 11/08/2005
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Avi Biopharma, Inc.
• Inventors: MOURICH, DAN V., WELLER, DWIGHT D., IVERSEN, PATRICK L.
• Granted Patent: US 8,501,704 B2
• Time in Patent Office: Days
• US Class Current: 424/184.100
• 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

A method and compound for suppressing an immune response in a mammalian subject, for the treatment or prevention of an autoimmune condition or transplantation rejection are disclosed. The compound is an antisense oligonucleotide analog compound having a targeting sequence complementary to a preprocessed CTLA-4 mRNA region identified by SEQ ID NO: 22 in SEQ ID NO: 1, spanning the splice junction between intron 1 and exon 2 of the preprocessed mRNA of the subject. The compound is 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 method of inducing immunological tolerance in a subject, for the prevention or treatment of an autoimmune condition, comprising

(a) administering to the subject, a pharmaceutically effective amount of an antisense compound composed of (i) 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′

-exocycklic carbon of an adjacent subunit and having a targeting sequence that contains at least 12 contiguous bases that are complementary to SEQ ID 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 human CTLA-4 mRNA, a heteroduplex structure characterized by a Tm of dissociation of at least 45°

C.,

(b) monitoring the subject by tests appropriate to the condition being treated, and

(c) repeating said administering until improvement in the disorder is observed.

2. The method of claim 1, wherein said administering is by a parenteral route, in an amount between 5-250 mg compound/dose.

3. The method of claim 2, wherein said administering is by intravenous or intraperitoneal administration.

4. The method of claim 1, wherein the oligonucleotide has one of the overlapping sequences identified as SEQ ID NOS:

4-6.

5. The method of claim 1, wherein the arginine-rich peptide in the compound administered in step (a) has one of the sequence identified by SEQ ID NOS:

15-21.

6. The method of claim 1, for use in delaying the onset of Type-1 diabetes in a subject, wherein monitoring the subject in step (b) includes measuring the blood sugar levels of the subject.

7. The method of claim 6, wherein said administering is by a parenteral route, in an amount between 5-250 mg compound/dose.

8. The method of claim 7, wherein said administering is by intravenous or intraperitoneal administration.

9. A composition for suppressing an immune response in a subject, for the treatment or prevention of an autoimmune condition, comprising

(a) a pharmaceutically effective amount of compound composed of (i) a morpholino antisense oligonucleotide antisense compound composed of (i) 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′

-exocycklic carbon of an adjacent subunit and having a targeting sequence that contains at least 12 contiguous bases that are complementary to SEQ ID 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 CTLA-4 mRNA, a heteroduplex structure characterized by a Tm of dissociation of at least 45°

C.; and

(b) a pharmaceutically acceptable carrier.

10. The composition of claim 9, wherein said morpholino subunits in the antisense oligonucleotide are joined by phosphorodiamidate linkages, in accordance with the structure:

11. The composition of claim 9, in which at least 2 and no more than half of the total number of intersubunit linkages in the antisense oligonucleotide are positively charged at physiological pH, and wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

12. The composition of claim 9, wherein the arginine-rich peptide in the compound has one of the sequences identified as SEQ ID NOS:

15-21.

13. The composition of claim 9, wherein the arginine-rich peptide in the compound has one of the sequences identified as SEQ ID NOS:

16.

14. The composition of claim 9, wherein the arginine-rich peptide in the compound has one of the sequences identified as SEQ ID NOS:

21.

15. The composition of claim 9, wherein the oligonucleotide is contained within includes one of the overlapping sequences identified as SEQ ID NOS:

4-6.

16. The composition of claim 14, wherein the oligonucleotide includes SEQ ID NO:

4.