IP Library Granted Patent US 8,912,316
Granted Patent B2
US 8,912,316 · App. 13/612,521 · Granted Dec 16, 2014

Compositions and methods for inhibiting expression of CD45 gene

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Quick Facts
Patent No.
US 8,912,316
App. No.
13/612,521
Granted
Dec 16, 2014
Kind
B2
Abstract

The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of the CD45 gene.

Claims (27)

1. A double-stranded ribonucleic acid (dsRNA), wherein said dsRNA comprises a sense strand and an antisense strand that are substantially complementary to each other, and wherein said antisense strand comprises a region that is substantially complementary to a corresponding region less than 30 nucleotides in length of an mRNA encoding CD45, and wherein said antisense strand comprises at least 15 contiguous nucleotides of SEQ ID NO:148, and wherein said dsRNA targets cleavage of the CD45 mRNA.

2. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing said CD45 gene, inhibits expression of said CD45 gene by at least 20% as compared to a control cell expressing said CD45 gene and not in contact with said dsRNA.

3. The dsRNA of claim 1 , wherein at least one strand of said dsRNA comprises at least one modified nucleotide.

4. The dsRNA of claim 3 , wherein said modified nucleotide is chosen from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.

5. The dsRNA of claim 3 , wherein said modified nucleotide is chosen from the group consisting of: a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

6. The dsRNA of claim 1 , wherein said sense strand and said antisense strand form a region of complementarity and said region of complementarity is at least 15 nucleotides in length.

7. The dsRNA of claim 6 , wherein said region of complementarity is 19-24 nucleotides in length.

8. The dsRNA of claim 1 , wherein said dsRNA, upon contact with a cell expressing CD45, inhibits expression of CD45 by at least 20% as measured in the P388D1 cell assay as compared to a control cell expressing CD45 and not in contact with said dsRNA.

9. A cell comprising the dsRNA of claim 1 .

10. A pharmaceutical composition comprising the dsRNA of claim 1 , and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition of claim 10 , wherein said dsRNA, upon contact with a cell expressing said CD45 gene, inhibits expression of said CD45 gene by at least 20%, as measured in a P388D1 cell assay as compared to a control cell expressing CD45 and not in contact with said dsRNA.

12. A method for inhibiting expression of a CD45 gene in a cell, the method comprising:

(a) introducing into the cell the dsRNA of claim 1 ; and

(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the CD45 gene, thereby inhibiting expression of the CD45 gene in the cell.

13. The method of claim 12 , wherein the cell produced in step (a) is maintained for a time sufficient to inhibit expression of CD45 by 20% as compared to a control cell expressing CD45 into which said dsRNA has not been introduced.

14. A vector for inhibiting the expression of the CD45 gene in a cell, said vector comprising a regulatory sequence operably linked to a nucleotide sequence that encodes at least one strand of the dsRNA of claim 1 .

15. A cell comprising the vector of claim 14 .

16. The dsRNA of claim 1 , wherein said sense strand comprises at least 15 contiguous nucleotides of the nucleotide sequence of SEQ ID NO: 147.

17. The dsRNA of claim 1 , wherein said antisense strand comprises SEQ ID NO: 148.

18. The dsRNA of claim 1 , wherein the sense strand comprises SEQ ID NO:147 and the antisense strand comprises SEQ ID NO:148.

19. The dsRNA of claim 1 , wherein the sense strand comprises SEQ ID NO:243 and the antisense strand comprises SEQ ID NO:244.

20. The dsRNA of claim 1 , wherein the sense strand consists of SEQ ID NO:1 (GAAuAAAAAccGuuAuGuuTsT) and the antisense strand consists of SEQ ID NO:2 (AAcAuAACGGUUUUuAUUCTsT) and each strand includes a 2′-O-methyl ribonucleotide as indicated by a lower case letter “c” or “u” and a phosphorothioate as indicated by a lower case letter “s”.

21. The dsRNA of claim 19 , wherein at least one strand comprises at least one 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, or a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.

22. A method for inhibiting expression of a CD45 gene in a cell, the method comprising:

(a) introducing into the cell the dsRNA of claim 20 ; and

(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the CD45 gene, thereby inhibiting expression of the CD45 gene in the cell.

23. A pharmaceutical composition comprising the dsRNA of claim 20 and a pharmaceutically acceptable carrier.

Assignments (5)
SECURITY INTEREST Recorded Oct 1, 2025
From: ALNYLAM PHARMACEUTICALS, INC.; SIRNA THERAPEUTICS, INC.
To: BANK OF AMERICA, N.A.
Reel/Frame 072996/0337 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 5, 2012
From: TAN, PAMELA
To: ALNYLAM EUROPE AG
Reel/Frame 029086/0763 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 5, 2012
From: ALNYLAM EUROPE AG
To: ALNYLAM PHARMACEUTICALS, INC.
Reel/Frame 029086/0777 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 5, 2012
From: DE FOUGEROLLES, ANTONIN; BORODOVSKY, ANNA; NOVOBRANTSEVA, TATIANA
To: ALNYLAM PHARMACEUTICALS, INC.
Reel/Frame 029086/0807 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 5, 2012
From: BAVARI, SINA; WARFIELD, KELLY LYN
To: UNITED STATES ARMY
Reel/Frame 029086/0825 →