Pharmaceutical compositions comprising antisense oligonucleotides and methods of using same

• US 9,050,373 B2
• Filed: 05/13/2010
• Issued: 06/09/2015
• Est. Priority Date: 05/13/2010
• Status: Active Grant

• Current Assignee: Charlotte Mecklenburg Hospital Authority
• Original Assignee: Charlotte Mecklenburg Hospital Authority
• Inventors: Lu, Qi Long, Wu, Bo
• Primary Examiner(s): Angell, Jon E
• Application Number:
  US12/800,376
• Publication Number:
  US 20110281787A1
• Time in Patent Office: 1,853 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61K 47/6455 Polycationic oligopeptides, polypeptides or polyamino acids, e.g. for complexing nucleic acids
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 39/02 Antidotes
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring

Abstract

The invention provides compositions and associated methods for the antisense treatment of genetic disorders, infections and various other medical conditions. In particular, embodiments of the present invention are directed to pharmaceutical compositions comprising a combination of an antisense oligonucleotide compound conjugated with a positively charged polymer (“ON-PCP”) and a negatively charged polymer. Pharmaceutical compositions in accordance with the present invention have demonstrated improved antisense efficiency and reductions in cell toxicity compared to compositions that contain an oligonucleotide compound conjugated with a positively charged polymer.

Claims

1. A pharmaceutical composition comprising a mixture of an antisense oligonucleotide conjugated with a positively charged polymer, wherein the antisense oligonucleotide comprises a morpholino oligomer;

and a negatively charged polymer, the composition when administered exhibits improved antisense efficiency and/or a reduction in cell toxicity in comparison to a composition containing the oligomer conjugated with a positively charged polymer.

2. The composition of claim 1, wherein the antisense oligonucleotide comprises a morpholino oligomer having from about 15 to 40 base pairing units (monomers).

3. The composition of claim 2, wherein the morpholino oligomer comprises a phosphorodiamidate-linked morpholino oligomer.

4. The composition of claim 1, wherein the positively charged polymer is a peptide.

5. The composition of claim 1, wherein the positively charged polymer is a cationic polymer.

6. The composition of claim 1, wherein the positively charged polymer is dendrimeric octaguanidine polymer.

7. The composition of claim 1, wherein the negatively charged polymer comprises an RNA or DNA strand having from about 2 to 50 base pairs.

8. The composition of claim 1, wherein the negatively charged polymer comprises a polysaccharide.

9. The composition of claim 1, wherein the negatively charged polymer comprises 2′

-O-methyl phosphorothioate oligonucleotide.

10. The composition of claim 1, wherein the negatively charged polymer comprises 2′

-O-methoxyethyl phosphorothioate oligonucleotide.

11. The composition of claim 1, wherein the negatively charged polymer is ionically bound to a positively charged polymer conjugated with the antisense oligonucleotide.

12. The composition of claim 1, wherein the ratio of the negatively charged polymer to the antisense oligomer conjugated with a positively charged polymer is from about 1:

100 to 100;

1.

13. The composition of claim 1, wherein the ratio of the negatively charged polymer to the antisense oligomer conjugated with a positively charged polymer is from about 0.2:

1 to 1;

5.

14. A pharmaceutical composition comprising

a phosphorodiamidate-linked morpholino oligomer conjugated with a positively charged polymer;

a negatively charged polymer; and

a carrier, the composition when administered exhibits improved antisense efficiency and/or a reduction in toxicity in comparison to a composition containing the oligomer conjugated with a positively charged polymer.

15. The pharmaceutical composition according to claim 14, wherein the antisense oligomer and the negatively charged polymer are both complimentary, near complimentary or non complimentary to a same target region of DNA or RNA.

16. The pharmaceutical composition according to claim 14, wherein the negatively charged polymer comprises 2′

-O-methyl phosphorothioate oligonucleotide or 2′

-O-methoxyethyl phosphorothioate oligonucleotide.

17. A method for treating a medical condition to a subject in need thereof comprising administering to a patient a therapeutically effective amount of a pharmaceutical composition comprising an antisense oligonucleotide conjugated with a positively charged polymer;

and a negatively charged polymer, the composition when administered exhibits improved antisense efficiency and/or reduced toxicity in comparison to a composition containing the oligomer conjugated with a positively charged polymer.

18. The method of claim 17, wherein the antisense oligonucleotide comprises phosphorodiamidate-linked morpholino oligomer or a PNA oligomer.

19. The method of claim 17, wherein the positively charged polymer is a peptide, cationic polymer, or a combination thereof.

20. The method of claim 17, wherein a solution containing the negatively charged polymer is mixed with a solution containing the antisense oligonucleotide conjugated with a positively charged polymer prior to the step of administering the pharmaceutical composition.

21. The method of claim 17, wherein the medical condition is a bacterial or viral infection.

22. The method of claim 17, wherein the medical condition is cancer.

23. The method of claim 17, wherein the medical condition is a muscle related disease.

24. The method of claim 17, wherein the medical condition is a muscular dystrophy.

25. The method of claim 17, wherein the pharmaceutical composition is administered at a dosage from about 0.5 mg to 300 mg/kg body weight.

26. A pharmaceutical composition comprising a mixture of an antisense oligonucleotide conjugated with a positively charged polymer, wherein the antisense oligonucleotide comprises a morpholino oligomer;

and a negatively charged polymer, the composition when administered exhibits improved antisense efficiency and/or reduced toxicity in comparison to a composition containing the oligomer conjugated with a positively charged polymer; and

wherein the negatively charged polymer comprises the same or similar sequence as the antisense oligonucleotide.

27. A pharmaceutical composition of claim 26, wherein the negatively charged polymer is selected from the group consisting of RNA, DNA, 2′

-O-methyl phosphorothioate oligonucleotides, 2′

-O-methoxy-ethyl phosphorothioate oligonucleotides, and combinations thereof.