Note: Claims are shown in the official language in which they were submitted.
CLAIMS:
1. An RNA molecule that down regulates the expression of a target mRNA,
the RNA molecules comprising a first strand that is complementary to a human
target
mRNA and a second strand complementary to the first strand, wherein the first
strand and
second strands can anneal to form a double-stranded region having from about
15 base
pairs to about 40 base pairs.
2. The RNA molecule of claim 1 wherein the first strand is from 19 to 24
nucleotides in length or from 25 to 29 nucleotides in length and is
complementary to a
human target gene selected from the group consisting of tumor necrosis factor
(TNF),
vascular endothelial growth factor (VEGF), vascular endothelial growth factor
receptor
(VEGFR), epidermal growth factor receptor (EGFR), erythroblastic leukemia
viral
oncogene homolog (ERBB), platelet derived growth factor (PDGF), platelet
derived
growth factor receptor (PDGFR), breakpoint cluster region (BCR)-abelson murine
leukemia viral oncogene homolog (ABL), steroid-5-alpha-reductase, alpha
polypeptide 1
(SRD5A1), steroid-5-alpha-reductase, alpha polypeptide 2 (SRD5A2),
phosphoinositide-
3-kinase, catalytic (PIK3C), mitogen-activated protein kinase (MAPK), p38 MAPK
family, hypoxia-inducible factor 1 alpha (HIF1A), protein kinase N3 (PKN3),
interleukin
17A (IL17A), interleukin 6(IL6), interleukin 18 (IL18), tumor necrosis factor
(ligand)
superfamily member 13b (TNFSF13B), mitogen-activated protein kinase 1(MAPK1),
v-
raf-1 murine leukemia viral oncogene homolog 1(RAF1), v-AKT murine thymoma
viral
oncogene (AKT), FK506 binding protein 12-rapamycin associated protein
1(FRAP1),
mitogen-activated protein kinase 2 (MAPK2), cyclin-dependent kinase 2 (CDK2),
ATP-
binding cassette, subfamily B, member 1(ABCB1), B-cell CLL/lymphoma 2 (BCL2),
angiopoietin 2 (ANGPT2), checkpoint kinase 1 homolog (CHEKI), insulin-like
growth
factor 1 receptor (IGF1R), signal transducer and activator of transcription 3
(STAT3),
matrix metalloproteinase (MMP), folate hydrolase (prostate-specific membrane
antigen) 1
(FOLH1), v-myc myelocytomatosis viral oncogene homolog (avian) (MYC),
telomerase
RNA component (TERC), telomerase reverse transcriptase (TERT), protein kinase
C,
alpha (PRKCA), RAS viral (v-ras) oncogene homolog (RAS), chemokine (C-X-C
motif)
ligand or receptor (CXC), Wingless-Type MMTV (Murine Mammary Tumor Virus)
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Integration Site (WNT), toll-like receptor (TLR), Fc fragment of IgE, low
affinity II,
receptor for (CD23) (FCER2), FOS gene, (FOS, FOSB, FOSL1, OR FOSL2),
hydroxysteroid (11-beta) dehydrogenase (HSD11B1), JUN gene (cJUN, JUNB, or
JUND),
thymidine phosphorylase (TYMP), early growth response (EGR), zeste homolog 2
(EZH2), cyclin D1 (CCND1), Fas (TNF receptor superfamily, member 6) (FAS),
proliferating cell nuclear antigen (PCNA), fibroblast growth factor 2 (FGF2),
tumor
growth factor-beta (TGF-.beta.), tumor growth factor-beta receptor (TGF-
.beta.R), tumor-
associated calcium signal transducer 1(TACSTD1), Mucin 1(MUC1), protein
tyrosine
phosphatase, non-receptor-11 (Noonan Syndrome 1) (PTPN11), neuregulin 1(NRG1),
membrane metallo-endopeptidase (MME), CD19 molecule (CD19), CD40 molecule, TNF
receptor superfamily member 5(CD40), apolipoprotein B (including Ag(x)
antigen)
(ApoB), synuclein, alpha (non A4 component of amyloid precursor) (SNCA),
silent
mating type information regulation 2 homolog (SIRT2), histone deacetylase
(HDAC),
membrane-spanning 4-domains, subfamily A, member 1(MS4A1), CD22 molecule
(CD22), diacylglycerol o-acyltransferase 1(DGAT1), diacylglycerol o-
acyltransferase 2
(DGAT2), CD3 molecule (CD3), proprotein convertase subtilisin-like kexin type
9
(PCSK9), MET (Mesenchymal epithelial transition factor) (c-Met proto-
oncogene),
catenin (cadherin-associated protein) (beta-catenin) (CTNNB1), inhition of DNA
binding
proteins (Inhibition of Differentiation Proteins, Dominant Negative Helix-Loop-
Helix
Protein) (ID), protein tyrosine phosphatase, non-receptor type 1(PTPN1), tie-1
(TIE1;
tyrosine kinase with immunoglobulin and EGF factor homology domains 1), tek
tyrosine
kinase (TEK), fibroblast growth factor receptor (FGFR), mitogen-activated
protein kinase
3 (MAPK3), survivin (BIRC5), and polo-like kinase family genes (PLK1).
3. The RNA molecule of claim 1 wherein the RNA molecule has at least one
blunt end.
4. The RNA molecule of claim 1 wherein the RNA molecule has at least one
3'-overhang.
5. The RNA molecule of claim 1 further comprising at least one acyclic
nucleomonomer.
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6. The RNA molecule of claim 5 wherein the acyclic nucleomonomer is
selected from the group consisting of
Image
wherein,
R is selected from the group consisting of hydrogen, a methyl group, C(1-
10) alkyl, cholesterol, naturally or non-naturally occurring amino acid,
sugar, vitamin,
flurophore, polyamine and fatty acid.
7. The RNA molecule of claim 6 wherein at least one acyclic nucleomonomer
is linked to the blunt end of the RNA molecule.
8. The RNA molecule of claim 6 at least one acyclic nucleomonomer is in the
double-stranded region of the RNA molecule.
9. A method for reducing the expression of a human target gene, comprising
administering an RNA molecule of claim 1 to a cell expressing the target gene,
wherein
the RNA molecule reduces expression of the target gene in the cell.
10. The method according to claim 9 wherein the cell is a human cell.
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11. A meroduplex ribonucleic acid (mdRNA) molecule that down regulates the
expression of a target mRNA, the mdRNA molecules comprising a first strand of
15 to 40
nucleotides in length that is complementary to a target mRNA and a second
strand and a
third strand that are each complementary to non-overlapping regions of the
first strand,
wherein the second strand and third strand can anneal with the first strand to
form at least
two double-stranded regions spaced apart by a nick or a gap.
12. The mdRNA molecule of claim 11 wherein the first strand is 15 to
25 nucleotides in length or 26 to 40 nucleotides in length.
13. The mdRNA molecule of claim 11 wherein the first strand is from 19 to 24
nucleotides in length or from 25 to 29 nucleotides in length and is
complementary to a
human target gene selected from the group consisting of tumor necrosis factor
(TNF),
vascular endothelial growth factor (VEGF), vascular endothelial growth factor
receptor
(VEGFR), epidermal growth factor receptor (EGFR), erythroblastic leukemia
viral
oncogene homolog (ERBB), platelet derived growth factor (PDGF), platelet
derived
growth factor receptor (PDGFR), breakpoint cluster region (BCR)-abelson murine
leukemia viral oncogene homolog (ABL), steroid-5-alpha-reductase, alpha
polypeptide 1
(SRD5A1), steroid-5-alpha-reductase, alpha polypeptide 2 (SRD5A2),
phosphoinositide-
3-kinase, catalytic (PIK3C), mitogen-activated protein kinase (MAPK), p38 MAPK
family, hypoxia-inducible factor 1 alpha (HIF1A), protein kinase N3 (PKN3),
interleukin
17A (IL17A), interleukin 6(IL6), interleukin 18 (IL18), tumor necrosis factor
(ligand)
superfamily member 13b (TNFSF13B), mitogen-activated protein kinase 1(MAPK1),
v-
raf-1 murine leukemia viral oncogene homolog 1(RAF1), v-AKT murine thymoma
viral
oncogene (AKT), FK506 binding protein 12-rapamycin associated protein
1(FRAP1),
mitogen-activated protein kinase 2 (MAPK2), cyclin-dependent kinase 2 (CDK2),
ATP-
binding cassette, subfamily B, member 1(ABCB1), B-cell CLL/lymphoma 2 (BCL2),
angiopoietin 2 (ANGPT2), checkpoint kinase 1 homolog (CHEK1), insulin-like
growth
factor 1 receptor (IGF1R), signal transducer and activator of transcription 3
(STAT3),
matrix metalloproteinase (MMP), folate hydrolase (prostate-specific membrane
antigen) 1
(FOLH1), v-myc myelocytomatosis viral oncogene homolog (avian) (MYC),
telomerase
RNA component (TERC), telomerase reverse transcriptase (TERT), protein kinase
C,
alpha (PRKCA), RAS viral (v-ras) oncogene homolog (RAS), chemokine (C-X-C
motif)
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ligand or receptor (CXC), Wingless-Type MMTV (Murine Mammary Tumor Virus)
Integration Site (WNT), toll-like receptor (TLR), Fc fragment of IgE, low
affinity II,
receptor for (CD23) (FCER2), FOS gene, (FOS, FOSB, FOSL1, OR FOSL2),
hydroxysteroid (11-beta) dehydrogenase (HSD11B1), JUN gene (cJUN, JUNB, or
JUND),
thymidine phosphorylase (TYMP), early growth response (EGR), zeste homolog 2
(EZH2), cyclin D1 (CCND1), Fas (TNF receptor superfamily, member 6) (FAS),
proliferating cell nuclear antigen (PCNA), fibroblast growth factor 2 (FGF2),
tumor
growth factor-beta (TGF-.beta.), tumor growth factor-beta receptor (TGF-
.beta.R), tumor-
associated calcium signal transducer 1(TACSTD1), Mucin 1(MUC1), protein
tyrosine
phosphatase, non-receptor-11 (Noonan Syndrome 1) (PTPN11), neuregulin 1(NRG1),
membrane metallo-endopeptidase (MME), CD19 molecule (CD19), CD40 molecule, TNF
receptor superfamily member 5(CD40), apolipoprotein B (including Ag(x)
antigen)
(ApoB), synuclein, alpha (non A4 component of amyloid precursor) (SNCA),
silent
mating type information regulation 2 homolog (SIRT2), histone deacetylase
(HDAC),
membrane-spanning 4-domains, subfamily A, member 1(MS4A1), CD22 molecule
(CD22), diacylglycerol o-acyltransferase 1(DGAT1), diacylglycerol o-
acyltransferase 2
(DGAT2), CD3 molecule (CD3), proprotein convertase subtilisin-like kexin type
9
(PCSK9), MET (Mesenchymal epithelial transition factor) (c-Met proto-
oncogene),
catenin (cadherin-associated protein) (beta-catenin) (CTNNB1), inhition of DNA
binding
proteins (Inhibition of Differentiation Proteins, Dominant Negative Helix-Loop-
Helix
Protein) (ID), protein tyrosine phosphatase, non-receptor type 1(PTPN1), tie-1
(TIE1;
tyrosine kinase with immunoglobulin and EGF factor homology domains 1), tek
tyrosine
kinase (TEK), fibroblast growth factor receptor (FGFR), mitogen-activated
protein kinase
3 (MAPK3), survivin (BIRC5), and polo-like kinase family genes (PLK1).
14. The mdRNA molecule of claim 11 wherein the gap comprises from 1 to 10
unpaired nucleotides.
15. The mdRNA molecule of claim 11 wherein the mdRNA molecule has at
least one blunt end.
16. The mdRNA molecule of claim 11 wherein the mdRNA molecule has at
least one 3'-overhang comprising one to four nucleotides that are not part of
the gap.
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17. The mdRNA molecule of claim 11 further comprising at least on acyclic
nucleomonomer.
18. The mdRNA molecule of claim 17 wherein the at least one acyclic
nucleomonomer selected from the group consisting of:
Image
wherein,
R is selected from the group consisting of hydrogen, methyl group, C(1-10)
alkyl, cholesterol, naturally or non-naturally occurring amino acid, sugar,
vitamin, flurophore, polyamine and fatty acid.
19. The mdRNA molecule of claim 18 wherein at least one acyclic
nucleomonomer is linked to the blunt end of the mdRNA molecule.
20. The mdRNA molecule of claim 18 at least one acyclic nucleomonomer is
in one of the double-stranded regions of the mdRNA molecule.
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21. A method for reducing the expression of a human target gene, comprising
administering an mdRNA molecule of claim 11 to a cell expressing the target
gene,
wherein the mdRNA molecule reduces expression of the target gene in the cell.
22. The method according to claim 21 wherein the cell is a human cell.
23. Use of an mdRNA molecule or dsRNA molecule as defined in any one of
the preceding claims for the manufacture of a medicament for use in the
therapy of cancer,
a metabolic disease or inflammatory disease.
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