Molecule for treating an inflammatory disorder

• US 8,802,645 B2
• Filed: 06/21/2012
• Issued: 08/12/2014
• Est. Priority Date: 12/24/2009
• Status: Active Grant

• Current Assignee: Academisch Ziekenhuis Leiden H.O.D.N. Lumc, BioMarin Technologies B.V.
• Original Assignee: Academisch Ziekenhuis Leiden H.O.D.N. Lumc, Prosensa Technologies B.V.
• Inventors: Van Ommen, Garrit-Jan Boudewijn, Aartsma-Rus, Annemieke, Van Deutekom, Judith Christina Theodora, De Kimpe, Josephus Johannes, Verbeek, Joseph Stephan, Ylmaz-Elis, Aliye Seda
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US13/529,640
• Publication Number:
  US 20120259002A1
• Time in Patent Office: 782 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/7115 Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
• CPC Class Codes: A61K 31/712 Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12N 2320/33 Alteration of splicing

Abstract

The invention provides two types of oligonucleotides for treating an inflammatory disorder: an oligonucleotide which is able of altering the splicing of a pre-mRNA encoding a C5 in order to decrease the amount of a C5a and an oligonucleotide which is able of altering the splicing of a pre-mRNA encoding a IL-1RAcP in order to increase the amount of a soluble IL-1RAcP. The invention further provides the use of said oligonucleotides for preventing or treating an inflammatory disorder.

Claims

1. An oligonucleotide which alters the splicing of a pre-mRNA encoding IL-1RAcP, wherein said oligonucleotide binds a part of exon 9 in the pre-mRNA encoding IL-1RAcP, wherein exon 9 comprises SEQ ID NO:

5 or 6, said oligonucleotide comprising a modification selected from the group consisting of;

a modification to increase resistance to RNAseH within a cell, a backbone modification and/or a sugar modification.

2. An oligonucleotide according to claim 1, wherein said oligonucleotide induces the skipping of exon 9 in the pre-mRNA encoding IL-1 RAcP.

3. An oligonucleotide according to claim 2, wherein said oligonucleotide inhibits the inclusion of exon 9 in of the pre-mRNA encoding IL-1 RAcP.

4. An oligonucleotide according to claim 1, wherein said oligonucleotide comprises a sequence which binds to at least a part of exon 9 in the pre-mRNA encoding IL-1RAcP wherein said part of exon 9 is a contiguous stretch comprising at least 8 nucleotides and wherein said oligonucleotide also binds to at least a part of a non-exon region of the pre-mRNA encoding IL-1RAcP wherein said part of the non-exon region is a contiguous stretch comprising at least 8 nucleotides.

5. An oligonucleotide according to claim 1, wherein said oligonucleotide comprises a sequence which also binds a splice site or an intronic sequence of the pre-mRNA encoding IL-1RAcP.

6. An oligonucleotide according to claim 4, wherein the contiguous stretch of exon 9 and/or the contiguous stretch of non-exon region comprises 8-50 nucleotides of exon 9 in the pre-mRNA encoding IL-1RAcP.

7. An oligonucleotide according to claim 1, wherein said oligonucleotide comprises or consists of a sequence selected from the group consisting of:

SEQ ID NOS;

25-41.

8. An oligonucleotide according to claim 1, wherein the oligonucleotide has a modified backbone and a modified sugar moiety wherein the oligonucleotide comprises one or more 2′

-O-methyl riboses and one or more phosphorothioate internucleoside linkages.

9. An oligonucleotide according to claim 8, wherein the oligonucleotide comprises a 2′

-O-methyl phosphorothioate internucleoside linkage and a locked nucleic acid monomer.

10. An oligonucleotide according to claim 1, wherein said oligonucleotide comprises at least one inosine and/or a base able to form a wobble base pair.

13. A composition comprising the oligonucleotide of claim 1.

14. A composition according to claim 13, further comprising a second oligonucleotide, wherein said second oligonucleotide alters the splicing of a pre-mRNA encoding C5 in order to decrease the amount of a C5a.

15. An oligonucleotide according to claim 6, wherein the contiguous stretch comprises 14-25 nucleotides of exon 9 in of the pre-mRNA encoding IL-1 RAcP.

16. A composition according to claim 13, wherein the composition is a pharmaceutical composition, said pharmaceutical composition comprising a pharmaceutically acceptable carrier, adjuvant, diluent and/or excipient.

18. The oligonucleotide of claim 1, wherein the modification to increase resistance to RNAseH within a cell is selected from the group consisting of:

a backbone modification, a sugar modification, and a base modification.

19. The oligonucleotide of claim 18, wherein the backbone modification is selected from the group consisting of:

a phosphorodithioate internucleoside linkage, a phosphorothioate internucleoside linkage, a chirally pure phosphorothioate internucleoside linkage, a methyl phosphonate internucleoside linkage, and a H-phosphonate internucleoside linkage.

20. The oligonucleotide of claim 18, wherein the sugar modification is selected from the group consisting of:

2′

-halide, 2′

-O-alkyl, 2′

-O-methyl, 2′

-F, 2′

-O-(2-methoxy)ethyl, 2′

-O-ethyl, 2′

-O-allyl, 2′

-O-butyryl, 2′

-O-propargyl, 2′

-O-(2-amino)ethyl, a locked nucleic acid monomer, and an ethylene-bridged nucleic acid monomer.

21. The oligonucleotide of claim 18, wherein the base modification is selected from the group consisting of:

5-halogenated uracil, 5-halogenated cytosine, 5-aminomethyl-uracil, 2,6-diaminopurine, 5-propargyl-cytosine, 5-propargyl-uracil, G-clamp and its derivatives, 5-methyl-cytosine and 5-methyl-uracil.

22. The oligonucleotide of claim 18, wherein the modification to increase resistance to RNAseH comprises a 2′

-O-methyl sugar modification and a phosphorothioate internucleoside linkage backbone modification.

23. The oligonucleotide of claim 18, wherein the modification to increase resistance to RNAse H comprises a 2′

-O-methyl sugar modification, a phosphorothioate internucleoside linkage backbone modification and/or a locked nucleic acid monomer sugar modification.

11. A method for preventing or treating an inflammatory disorder in an individual, the method comprising administering to said individual an effective amount of an oligonucleotide wherein said oligonucleotide alters the splicing of a pre-mRNA encoding IL-1RAcP, binds a part of exon 9 of the IL-1RAcP pre-mRNA, wherein exon 9 comprises SEQ ID NO:

5 or 6, said oligonucleotide comprising a modification selected from the group consisting of;

a modification to increase resistance to RNAseH within a cell, a backbone modification, and/or a sugar modification.

12. The method of claim 11, wherein said disorder comprises rheumatoid arthritis (RA), juvenile rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, inflammatory bowel disease including Crohn'"'"'s disease or ulcerative colitis, hepatitis, sepsis, alcoholic liver disease, or non-alcoholic steatosis.

17. An oligonucleotide which alters the splicing of a pre-mRNA encoding IL-1RAcP, wherein said oligonucleotide binds a part of a non-exon region of the pre-mRNA encoding IL-1RAcP, wherein exon 9 in the pre-mRNA encoding IL-1RAcP comprises SEQ ID NO:

5 or 6;

wherein said oligonucleotide induces the skipping of exon 9 in the pre-mRNA encoding IL-1RAcP, and wherein said oligonucleotide comprises a modification selected from the group consisting of;

a modification to increase resistance to RNAseH within a cell, a backbone modification and/or a sugar modification.