Pharmaceutical composition for treatment of Duchenne muscular dystrophy

• US 20020055481A1
• Filed: 08/16/2001
• Published: 05/09/2002
• Est. Priority Date: 08/25/2000
• Status: Active Grant

• Current Assignee: JCR Pharmaceuticals Co., Ltd., Masafumi Matsuo
• Original Assignee: JCR Pharmaceuticals Co., Ltd.
• Inventors: Matsuo, Masafumi, Kamei, Shoichiro
• Granted Patent: US 6,727,355 B2
• Time in Patent Office: Days
• US Class Current: 514/44
• 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 47/02 Inorganic compounds
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C07K 14/4708 Duchenne dystrophy

Abstract

The invention provides an isolated and purified DNA set forth as SEQ ID NO:15 in the Sequence Listing and an antisense oligonucleotide complementary to the DNA. The DNA represents the splicing enhancer sequence (SES) in exon 45 of human dystrophin gene, and serves as a template in preparation of the antisense oligonucleotide, which is used to induce exon 45 skipping in certain group of patient with Duchenne muscular dystrophy to restore the reading frame of dystrophin mRNA.

Claims

1. An isolated and purified oligonucleotide selected from the group consisting of a DNA comprising the nucleotide sequence set forth as SEQ ID NO:

15 in the Sequence Listing and an RNA comprising the nucleotide sequence complementary to the nucleotide sequence that in turn is complementary to the nucleotide sequence set forth as SEQ ID NO;

15 in the Sequence Listing.

2. An isolated and purified antisense oligonucleotide comprising the nucleotide sequence complementary to the nucleotide sequence set forth as SEQ ID NO:

15 in the Sequence Listing.

3. The antisense oligonucleotide of claim 2 selected from the group consisting of a DNA or phosphorothioate DNA comprising the nucleotide sequence set forth as SEQ ID NO:

19 in the Sequence Listing.

4. Use of the antisense oligonucleotide of claim 2 or 3 for the manufacture of a pharmaceutical composition for treatment of Duchenne muscular dystrophy caused by an alteration in the number of the nucleotides in the nucleotide sequences encoding one or more exons adjacent to exon 45 in human dystrophin mRNA due to deletion of one or more nucleotides from the normal nucleotide sequences, wherein the net of the alteration in the number of the nucleotides is expressed as a loss of (3×

N+1) nucleotides, wherein N is zero or a natural number.

5. A pharmaceutical composition for treatment of Duchenne muscular dystrophy which composition comprises the antisense oligonucleotide of claim 2 or 3 in a pharmaceutically acceptable injectable medium.

6. The pharmaceutical composition of claim 5 comprising 0.05-5 μ

mol/ml of the antisense oligonucleotide.

7. The pharmaceutical composition of claim 5 or 6 comprising 0.02-10 w/v % of at least one carbohydrate or polyalcohol, and 0.01-0.4 w/v % of at least one pharmaceutically acceptable surfactant.

8. The pharmaceutical composition of one of claims 5 to 7 comprising 0.03-0.09 M of at least one pharmaceutically acceptable neutral salt.

9. The pharmaceutical composition of claim 8 wherein the neutral salt is selected from the group consisting of sodium chloride, potassium chloride, and calcium chloride.

10. The pharmaceutical composition of one of claims 5 to 9 comprising 0.002-0.05 M of a pharmaceutically acceptable buffering agent.

11. The pharmaceutical composition of claim 10 wherein the buffering agent is selected from the group consisting of sodium citrate, sodium glycinate, sodium phosphate, and tris(hydroxymethyl)aminomethane.

12. The pharmaceutical composition claim 7 wherein the carbohydrate is selected from the group consisting of monosaccharides and disaccharides.

13. The pharmaceutical composition of claim 7 or 12 wherein the carbohydrate or polyalcohol is selected from the group consisting of glucose, galactose, mannose, lactose, maltose, mannitol, and sorbitol.

14. The pharmaceutical composition of claim 7 or 12 wherein the surfactant is selected from the group consisting of polyoxyethylene sorbitan mono- to tri-ester, alkyl phenyl polyoxyethylene, sodium taurocholate, sodium cholate, and a polyalcohol ester.

15. The pharmaceutical composition of claim 14 wherein the polyoxyethylene sorbitan ester is an ester selected from the group consisting of oleate, laurate, stearate, and palmitate.

16. The pharmaceutical composition of one of claims 5 to 15 in which the antisense oligonucleotide is provided in a lyophilized form for reconstitution prior to use for a patient.

18. The method of claim 17 wherein the antisense oligonucleotide is selected from the group consisting of a DNA or phosphorothioate DNA comprising the nucleotide sequence set forth as SEQ ID NO:

15 in the Sequence Listing.

17. A method for treatment of a human patient with Duchenne muscular dystrophy caused by an alteration in the number of the nucleotides in the nucleotide sequences encoding one or more exons adjacent to exon 45 in human dystrophin mRNA due to deletion of one or more nucleotides from the normal nucleotide sequences, wherein the net of the alteration in the number of the nucleotides is expressed as a loss of (3×

N+1) nucleotides, wherein N is zero or a natural number, wherein the method comprises administering to the patient an therapeutically effective amount of an isolated and purified antisense oligonucleotide comprising the nucleotide sequence complementary to the nucleotide sequence set forth as SEQ ID NO;

15 in the Sequence Listing in a pharmaceutically acceptable injectable medium.