Modulation of CD40 expression

• US RE47,320 E1
• Filed: 12/21/2016
• Issued: 03/26/2019
• Est. Priority Date: 11/20/2007
• Status: Active Grant

• Current Assignee: Ionis Pharmaceuticals, Inc.
• Original Assignee: Ionis Pharmaceuticals, Inc.
• Inventors: Bennett, C. Frank, Cowsert, Lex M., Freier, Susan M.
• Primary Examiner(s): Ponnaluri, Padmashri
• Application Number:
  US15/386,791
• Time in Patent Office: 825 Days
• Field of Search: 536 245, 536 231, 536 253
• 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 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3231 having an additional ring, e.g. LNA, ENA
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3525 MOE, methoxyethoxy

Abstract

Disclosed herein are antisense compounds and methods for decreasing CD40. Examples of disease conditions that can be ameliorated with the administration of antisense compounds targeted to CD40 include hyperproliferative disorders, graft versus host disease (GVHD), graft rejection, asthma, airway hyperresponsiveness, chronic obstructive pulmonary disease (COPD), multiple sclerosis (MS), systemic lupus erythematosus (SLE), and certain forms of arthritis.

Claims

1. A modified antisense compound 12 to 30 nucleobases in length and having a nucleobase sequence that is at least 90% complementary to an equal length portion of the human CD40 gene but not to other sequences throughout the human genome, selected from the following regions of SEQ ID NO:

4;

(a) positions 11250-12685, corresponding to intron 6 11801-12591;

(b) positions 2943-6367, corresponding to intron 1;

(c) positions 6447-6780, corresponding to intron 2;

(d) positions 6907-7157, corresponding to intron 3;

(e) positions 7305-7673, corresponding to intron 4;

(f) positions 7768-11187, corresponding to intron 5;

(g) positions 12773-12877, corresponding to intron 7;

(h) positions 12907-13429, corresponding to intron 8; and

(i) positions 13662-16001, which forms part of exon 9 or a region 3′

to exon 9.

2. The antisense compound of claim 1, wherein the nucleobase sequence is at least 90% complementary to an equal length portion of positions 12527-12685 of SEQ ID NO:

4.

3. The antisense compound of claim 2 1, having a nucleobase sequence comprising at least 8 contiguous nucleobases of the nucleobase sequence of SEQ ID NO:

208, wherein the nucleobase sequence of the compound is at least 95% complementary to the sequence shown in SEQ ID NO;

4.

5. The antisense compound of claim 1, wherein said antisense compound is an antisense oligonucleotide.

6. The antisense compound of claim 5, wherein at least one internucleoside linkage is a modified internucleoside linkage.

7. The antisense compound of claim 6, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

8. The antisense compound of claim 5, wherein at least one nucleoside comprises a modified sugar.

9. The antisense compound of claim 8, wherein at least one modified sugar is a bicyclic sugar.

10. The antisense compound of claim 9, wherein the at least one bicyclic sugar comprises a 4′

-CH(CH₃)—

O-2′

bridge.

11. The antisense compound of claim 8, wherein at least one modified sugar comprises a 2′

-O-methoxyethyl.

12. The antisense compound of claim 1, wherein at least one said nucleobase is a modified nucleobase.

13. The antisense compound of claim 12, wherein the modified nucleobase is a 5-methylcytosine.

14. The antisense compound of claim 1, wherein the compound is an oligonucleotide comprising:

a gap segment consisting of linked deoxynucleosides;

a 5′

wing segment consisting of linked nucleosides;

a 3′

wing segment consisting of linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.

15. The antisense compound of claim 14, wherein the oligonucleotide comprises:

a gap segment consisting of ten linked deoxynucleosides;

a 5′

wing segment consisting of five linked nucleosides;

a 3′

wing segment consisting of five linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar;

and wherein each internucleoside linkage of said antisense compound is a phosphorothioate linkage.

16. The antisense compound of claim 14, wherein the oligonucleotide comprises:

a gap segment consisting of fifteen linked deoxynucleosides;

a 5′

wing segment consisting of two linked nucleosides;

a 3′

wing segment consisting of three linked nucleosides;

wherein the gap segment is positioned between the 5′

wing segment and the 3′

wing segment, wherein each nucleoside of each wing segment comprises a 2′

-O-methoxyethyl sugar;

and wherein each internucleoside linkage of said antisense compound is a phosphorothioate linkage.

17. The antisense compound of claim 15 or 16, wherein every cytosine is a 5-methylcytosine.

19. A composition comprising an antisense compound of claim 1 or 18 or a salt thereof and a pharmaceutically acceptable carrier or diluent.

20. A method comprising administering to an animal an antisense compound of claim 1 or an oligonucleotide of claim 18.

21. The antisense compound of claim 1, wherein said antisense compound is 15, 16, 17, 18, 19, 20 or 21 nucleobases in length.

22. The antisense compound of claim 21, wherein the nucleobase sequence of the compound is at least 95% complementary to the sequence shown in SEQ ID NO:

4.

23. The antisense compound of claim 21, wherein the nucleobase sequence of the compound is 100% complementary to the sequence shown in SEQ ID NO:

4.

4. A modified antisense compound 20 nucleobases in length and consisting of the nucleobase sequence of SEQ ID NO:

208.

18. An antisense oligonucleotide 20 nucleobases in length having the sequence of nucleobases as set forth in SEQ ID NO:

208, wherein each cytosine is a 5-methylcytosine, each internucleoside linkage is a phosphorothioate linkage, nucleotides 1-5 and 16-20 are 2′

-O-methoxyethyl nucleotides, and nucleotides 6-15 are 2′

-deoxynucleotides.