METHODS AND COMPOSITIONS FOR TREATING NEUROLOGICAL DISEASE

• US 20100267810A1
• Filed: 06/25/2010
• Published: 10/21/2010
• Est. Priority Date: 08/18/2005
• Status: Abandoned Application

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: ZAMORE, PHILLIP D., ARONIN, NEIL
• Application Number:
  US12/823,917
• Publication Number:
  US 20100267810A1
• Time in Patent Office: Days
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2320/32 Special delivery means, e.g. tissue-specific
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/136 Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA

Abstract

This invention relates to methods and compositions for treating neurological disease, and more particularly to methods of delivering iRNA agents to neural cells for the treatment of neurological diseases.

Claims

1. A method of downregulating expression of a target gene in a neural cell distal to the site of administration, the method comprising contacting an iRNA agent with the neural cell for a time sufficient to allow uptake of the iRNA agent into the cell, wherein (i) the iRNA agent comprises a sense and an antisense strand that form an RNA duplex, and (ii) the sequence of the antisense strand of the iRNA agent comprises a nucleotide sequence sufficiently complementary to a target sequence of about 18 to 25 nucleotides of an RNA expressed from the target gene.

3. The method of claim 1 or 2, wherein the cells are contacted for a time sufficient to allow axonal transport of said iRNA.

4. The method of claim 1, wherein the iRNA agent comprises a lipophilic moiety.

5. The method of claim 4, wherein the lipophilic moiety is a cholesterol.

6. The method of claim 4, wherein the lipophilic moiety is conjugated to the sense strand.

7. The method of claim 4, wherein the lipophilic moiety is conjugated to the 3′

end of the sense strand.

2. A method of downregulating expression of a target gene in a neural cell, the method comprising contacting an iRNA agent with the neural cell for a time sufficient to allow uptake of the iRNA agent into the cell, wherein (i) the iRNA agent comprises a sense and an antisense strand that form an RNA duplex, (ii) the iRNA agent comprises a lipophilic moiety, and (iii) the sequence of the antisense strand of the iRNA agent comprises a nucleotide sequence sufficiently complementary to a target sequence of about 18 to 25 nucleotides of an RNA expressed from the target gene.

8-12. -12. (canceled)

13. A method of treating a human comprising identifying a human having or at risk for developing a neurological disorder, the method comprising administering to the human an iRNA agent that targets a gene expressed in a neural cell distal to the site of administration, wherein the expression of the gene is associated with symptoms of the neurological disorder, and wherein (i) the iRNA agent comprises a sense and an antisense strand that form an RNA duplex, and (ii) the antisense strand of the iRNA agent comprises a nucleotide sequence sufficiently complementary to a target sequence of about 18 to 25 nucleotides of an RNA expressed from the target gene.

15. The method of claim 13, wherein the iRNA agent comprises a lipophilic moiety.

16. The method of claim 15, wherein the lipophilic moiety is a cholesterol.

17. The method of claim 15, wherein the lipophilic moiety is conjugated to the sense strand.

18. The method of claim 15, wherein the lipophilic moiety is conjugated to the 3′

end of the sense strand.

19. The method of claim 15, wherein the iRNA agent further comprises a phosphorothioate or a 2′

-OMe modification.

14. A method of treating a human comprising identifying a human having or at risk for developing a neurological disorder, and administering to the human an iRNA agent that targets a gene expressed in a neural cell, wherein the expression of the gene is associated with symptoms of the neurological disorder, and wherein (i) the iRNA agent comprises a sense and an antisense strand that form an RNA duplex, (ii) the iRNA agent comprises a lipophilic moiety, and (iii) the antisense strand of the iRNA agent comprises a nucleotide sequence sufficiently complementary to a target sequence of about 18 to 25 nucleotides of an RNA expressed from the target gene.

20-33. -33. (canceled)

34. A method of reducing the amount of huntingtin (htt) RNA in a neural cell of a subject, comprising:

contacting the neural cell with an iRNA agent, wherein said neural cell is distal to the site of action and the iRNA agent comprises a sense and an antisense strand, wherein the sense and the antisense strands form an RNA duplex, wherein the antisense strand comprises a nucleotide sequence that differs by no more than four nucleotides from an antisense sequence listed in Table 1.

35. The method of claim 34, wherein the iRNA agent further comprises a lipophilic moiety.

36. The method of claim 35, wherein the cells are contacted for a time sufficient to allow axonal transport of said iRNA.

37. The method of claim 36, wherein the iRNA agent further comprises a phosphorothioate or a 2′

-OMe modification.

38-42. -42. (canceled)

43. An isolated iRNA agent comprising a sense and an antisense strand, wherein the sense and the antisense strands form an RNA duplex, wherein the antisense strand comprises a nucleotide sequence that differs by no more than four nucleotides from an antisense sequence listed in Table 1, and wherein the iRNA agent comprises a lipophilic moiety.

44. The iRNA agent of claim 43, wherein the lipophilic moiety is a cholesterol molecule.

45. The iRNA agent of claim 43, wherein the lipophilic moiety is attached to the sense strand.

46. The iRNA agent of claim 43, wherein the lipophilic moiety is attached to the 3′

end of the sense strand.

47-51. -51. (canceled)

52. A pharmaceutical composition, comprising:

(i) an iRNA agent comprising a sense and an antisense strand, wherein the sense and the antisense strands form an RNA duplex, wherein the antisense strand comprises a nucleotide sequence that differs by no more than four nucleotides from an antisense sequence listed in Table 1, and wherein the iRNA agent comprises a lipophilic moiety; and

(ii) a pharmaceutically acceptable carrier.

53-56. -56. (canceled)

57. A method of evaluating an iRNA agent for enhanced uptake into neural cells comprising:

providing a candidate iRNA agent conjugated to a lipophilic agent, wherein the iRNA agent is in a solution that does not contain a transfection reagent, contacting the iRNA agent with a neural cell for a time sufficient for uptake into the neural cell and determining if the iRNA agent is taken up by the neural cell.

58-66. -66. (canceled)