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1. WO2006109196 - MODELES DE SOURIS TRANSGENIQUES DU VIRUS DE L'HEPATITE C (VHC) ET IDENTIFICATION DES THERAPIES POUR LE VHC

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WHAT IS CLAIMED IS:
1. A genetically modified mouse comprising a reduced sensitivity to TNF-alpha, an increased amount of cytoplasmic NFkB, a decreased amount of caspase 3, and a decreased amount of DNA fragmentation in liver cells, as compared to a wild-type mouse of the same variety, wherein said mouse further comprises an exogenous nucleic acid construct that comprises a promoter operably linked to a gene encoding a hepatitis C virus (HCV) nonstructural protein 3 (NS3).
2. The genetically modified mouse of Claim 1, further comprising a gene encoding a HCV nonstructural protein 4A (NS4A).
3. The genetically modified mouse of Claim 1 or 2, with the proviso that said genetically modified mouse does not comprise an HCV core protein, structural protein El (El), structural protein E2 (E2), nonstructural protein 2 (NS2), nonstructural protein 4B (NS4B), nonstructural protein 5A (NS5A), or nonstructural protein 5B (NS5B).
4. The genetically modified mouse of Claim 1, 2, or 3, wherein said promoter is a mouse major urinary promoter (MUP).
5. The genetically modified mouse of Claim 1, 2, 3, or 4, wherein said exogenous nucleic acid construct further comprises a nucleic acid encoding a NS3 protease cleavage site.
6. The genetically modified mouse of Claim 5, wherein said nucleic acid encoding said NS3 protease cleavage site is joined to said NS3 gene.
7. The genetically modified mouse of Claim 1, 2, 3, 4, 5, or 6, wherein said exogenous nucleic acid construct further comprises a nucleic acid encoding a secretory signal.
8. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, or 7, wherein said exogenous nucleic acid construct further comprises a nucleic acid encoding a signal molecule.
9. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, or 8, wherein said NS3 gene is codon optimized.
10. The genetically modified mouse of Claim 2, 3, 4, 5, 6, 7, 8, or 9, wherein said NS4A gene is codon optimized.
11. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, wherein said genetically modified mouse has an increased sensitivity to TNF-alpha after contact with 1) an inhibitor of a p38 MAP kinase or 2) an inhibitor of NF-kB, as compared to said genetically modified mouse that has not been contacted with said inhibitor.
12. The genetically modified mouse of Claim 11, wherein said inhibitor is SB203580.
13. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, wherein said genetically modified mouse comprises at-least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10.0 micrograms NS3 protein per gram of liver tissue.
14. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, wherein said genetically modified mouse comprises more than 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.1, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10.0 micrograms NS3 protein per gram of liver tissue.
15. The genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, wherein said genetically modified mouse comprises between about 0.1 - 1.0, 0.5 - 1.5, 1.0 - 2.0, 1.5 - 2.5, 2.0 - 3.0, 2.5 - 3.5, 3.0 - 4.0, 3.5 - 4.5, 4.0 - 5.0, 4.5 - 5.5, 5.0 - 6.0, 5.5 - 6.5, 6.0 - 7.0, 6.5 - 7.5, 7.0 - 8.0, 7.7 - 8.5, 8.0 - 9.0, 8.5 - 9.5, 9.0 - 10.0 micrograms NS3 protein per gram of liver tissue.
16. A method of using a genetically modified mouse to identify a compound that inhibits HCV replication comprising:
providing the genetically modified mouse of Claim 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15;
contacting said genetically modified mouse with a compound; and
analyzing the expression of NS3, NS3 protease activity, the sensitivity of said genetically modified mouse to TNF-alpha, the amount of NFkB in a cell, the amount of active caspase 3 in a cell, or the amout of DNA fragmentation in a liver cell after contact with said compound, whereby said compound that inhibits HCV replication is identified by the ability of said compound to inhibit expression of NS3, inhibit protease activity of NS3, restore TNF-alpha sensitivity, decrease the amount of NFkB in a cell, increase the amount of active caspase 3 in a cell, or increase the amount of DNA fragmentation in a liver cell in said genetically modified mouse.
17. A pharmaceutical or dietary supplement comprising the compound identified by the method of Claim 16.
18. The pharmaceutical or dietary supplement of Claim 17, wherein said compound is a p38 MAP kinase inhibitor and/or a NF-kB inhibitor.
19. The pharmaceutical or dietary supplement of Claim 18, wherein said compound is SB203580 or SB202190.
20. A method of inhibiting HCV infection comprising:
identifying a subject infected with HCV; and
providing said subject a composition comprising a p38 MAP kinase inhibitor and/or a NF-kB inhibitor.
21. The method of Claim 20, wherein said inhibitor is selected from the group consisting of SB 203580, SB 202190, KN62, U0126, PD 98059, Wortmannin, rapamycin, Ro 31-8220, Bis-1, Go 6976, UCNOl, Indirubin-3'-monoxime, kenpaullone, alsterpaullone, PPl, PP2, SU 6656, SP 600125, ML-9, PD 169316, p38 MAP Kinase inhibitor, SB202190 hydrochloride, SB 203580 hydrochloride, SB 203580 Iodo, SC68376, SKF-86002, ZM 3363772, Anti-p38/HOG-l antibody, and anti-p38 MAP Kinase antibody.
22. The method of Claim 20, wherein said inhibitor is SB203580 or SB202190.
23. The method of Claim 20, wherein said inhibitor is a suppressor of cytokine signaling- 1 (SOCSl) inhibitor.
24. The method of Claim 20, 21, 22, or 23, further comprising measuring the inhibition of HCV in said subject.
25. Use of a p38 MAP kinase inhibitor for the preparation of a medicament for the treatment or prevention of HCV infection.
26. The use of Claim 25, wherein said p38 inhibitor is SB203580 or SB202190.

27. An isolated totipotent mouse cell comprising an exogenous nucleic acid construct that comprises a mouse major urinary promoter (MUP) operably linked to a gene encoding an hepatitis C virus (HCV) nonstructural protein 3 (NS3).
28. The isolated totipotent mouse cell of Claim 27, further comprising nonstructural protein 4A (NS4A).
29. The isolated totipotent mouse cell of Claim 27, or 28, with the proviso that said isolated totipotent cell does not comprise an HCV core protein, structural protein El (El), structural protein E2 (E2), nonstructural protein 2 (NS2), nonstructural protein 4B (NS4B), nonstructural protein 5A (NS5A), or nonstructural protein 5B (NS5B).
30. Use of a NF-kB inhibitor for the preparation of a medicament for the treatment or prevention of HCV infection.
31. The use of Claim 30, further involving a p38 inhibitor in the preparation of the medicament.
32. The use of Claim 30, wherein said NF-kB inhibitor is selected from the group consisting of: Withaferin A; Sulfasalazine; PPM-18 (2-Benzoylamino-l,4-naphthoquinone); Oridonin; Parthenolide; NF-kB Activation Inhibitor; NF-kB SN50 (AAVALLPAVLLALLAPVQRKRQKLMP SEQ ID NO: 74), Cell-Permeable Inhibitor Peptide; NF-kB Activation Inhibitor; NEMO-Binding Domain Binding Peptide (DRQIKIWFQNRRMKWKKTALDWSWLQTE SEQ ID NO: 75); Kamebakaurin; Acetyl-11-keto-b-Boswellic Acid; Andrographolide; BAY 11-7082 ((E)3-[(4-Methylρhenyl)sulfonyl]-2-propenenitrile); BAY 11-7085 ((E)3-[(4-f-Butylphenyl)sulfonyl]-2-propenenitrile); CAPE (Caffeic Acid Phenethyl Ester); (E)-Capsaicin; Evodiamine; Gliotoxin; Helenalin; Hypoestoxide; Cell-Permeable; IkB Kinase Inhibitor Peptide, Cell-Permeable; Isohelenin; IKK-2 Inhibitor; IKK-2 Inhibitor VI; IKK-2 Inhibitor V; IKK-2 Inhibitor IV; IKIC Inhibitor III, BMS-345541; TIRAP Peptide (ToIl-interleukin 1 Receptor (TIR) domain-containing Adapter Protein Peptide); IKK Inhibitor II, Wedelolactone; IkB Kinase Inhibitor Peptide; Gliotoxin; an anti-inflamatory; aspirin; an NF-kB translocation inhibitor; an NF-IcB acetylation inhibitor; an inhibitor of an activator of NF-kB; an activator of an inhibitor of NF-kB; an HcB; an IkB-alpha; IkB-beta; IlcB-gamma; IkB-epsilon; Bcl-3; plOO; plO5; a non-steroidal anti inflammatory drug; and some combination thereof.
33. The use of Claim 32, wherein said non-steroidal anti inflammatory drug (NSAID) is selected from the group consisting of: ibuprofen (2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2,4 diamino 6-pteridinyl- methyl]methylamήio]benzoyl)-L-glutamic acid); aspirin (acetylsalicylic acid); salicylic acid; diphenhydramine (2-(diphenylmethoxy)-NN-dimethylethylamine hydrochloride); naproxen (2-naphthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (-)); phenylbutazone (4-butyl-l ,2-diphenyl-3,5-pyrazolidinedione); sulindac-(2)-5-fuoro-2-methyl-l-[[p-(methylsulfϊnyl)phenyl]methylene-]-lH- indene-3 -acetic acid; diflunisal (2',4',-difluoro-4-hydroxy-3-biphenylcarboxylic acid; piroxicam (4-hydroxy-2-methyl-N-2-pyridinyl-2H-l,2-benzothiazine-2-carboxamide 1,1-dioxide, an oxicam; indomethacin (1 -(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3-acetic acid); meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodium salt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid; tolmetin sodium (sodium l-methyl-5-(4-methylbenzoyl-lH-pyrrole-2-acetate dihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneatic acid, monosodium salt); hydroxychloroquine sulphate (2-{[4-[(7-chloro-4-quinolyl) amino]pentyl]ethylamino}ethanol sulfate (1:1); penicillamine (3-mercapto-D-valine); flurbiprofen ([l,l-biphenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+-.)); cetodolac (l-8-diethyl-13,4,9, tetra hydropyrano-[3-4-13]indole-l-acetic acid; mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydramine hydrochloride (2-diphenyl methoxy-N, N-di-methylethamine hydrochloride).
34. A method of inhibiting HCV infection comprising:
identifying a subject infected with HCV; and
providing said subject a composition comprising a NF-kB inhibitor.
35. The method of Claim 33, further comprising providing said subject a second composition comprising a p38 inhibitor.
36. The method of Claim 33, wherein said NF-kB and p38 inhibitors are a same composition.
37. The method of Claim 33, wherein said NF-kB inhibitor is selected from the group consisting of: Withaferin A; Sulfasalazine; PPM-18 (2-Benzoylamino-l,4-naphthoquinone); Oridonin; Parthenolide; NF-kB Activation Inhibitor; NF-kB SN50 (AAVALLPAVLLALLAPVQRKRQKLMP SEQ ID NO: 74), Cell-Permeable Inhibitor Peptide; NF-kB Activation Inhibitor; NEMO-Binding Domain Binding Peptide (DRQIKIWFQNRRMKWKKTALDWSWLQTE SEQ ID NO: 75); Kamebakaurin; Acetyl- 11-keto-b-Boswellic Acid; Andrographolide; BAY 11-7082 ((E)3-[(4-Methylphenyl)sulfonyl]-2-propenenitrile); BAY 11-7085 ((E)3-[(4-f-Butylphenyl)sulfonyl]-2-propenenitrile); CAPE (Caffeic Acid Phenethyl Ester); (E)-Capsaicin; Evodiamine; Gliotoxin; Helenalin; Hypoestoxide; Cell-Permeable; IkB Kinase Inhibitor Peptide, Cell-Permeable; Isohelenin; IKK-2 Inhibitor; IKK-2 Inhibitor VI; IKK-2 Inhibitor V; IKK-2 Inhibitor IV; IKK Inhibitor III, BMS-345541; TIRAP Peptide (ToIl-interleukin 1 Receptor (TIR) domain-containing Adapter Protein Peptide); IKK Inhibitor II, Wedelolactone; IkB Kinase Inhibitor Peptide; Gliotoxin; an anti-inflamatory; aspirin; an NF-kB translocation inhibitor; an NF-kB acetylation inhibitor; an inhibitor of an activator of NF-kB; an activator of an inhibitor of NF-kB; an IkB; an IkB-alpha; IkB-beta; IkB-gamma; IkB-epsilon; Bcl-3; plOO; plO5, non-steroidal anti inflammatory drug (NSAID); and some combination thereof.
38. The method of Claim 37, wherein said non-steroidal anti inflammatory drug (NSAID) is selected from the group consisting of: ibuprofen (2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2,4 diamino 6-pteridinyl-methyl]methylamino]benzoyl)-L-glutamic acid); aspirin (acetylsalicylic acid); salicylic acid; diphenhydramine (2-(diphenylmethoxy)-NN-dimethylethylamine hydrochloride); naproxen (2-naρhthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (-)); phenylbutazone (4-butyl-l,2-diphenyl-3,5-pyrazolidinedione); sulindac-(2)-5-fuoro-2-methyl-l-[[p-(methylsulfinyl)phenyl]methylene-]-lH-indene-3-acetic acid; diflunisal (2',4',-difluoro-4-hydroxy-3-biphenylcarboxylic acid; piroxicam (4-hydroxy-2-methyl-N-2-pyridinyl-2H-l,2-benzothiazine-2-carboxamide 1,1-dioxide, an oxicam; indomethacin ( 1 -(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3 -acetic acid) ; meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodium salt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid; tolmetin sodium (sodium l-methyl-5-(4-methylbenzoyl-lH-pyrrole-2-acetate dihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneatic acid, monosodium salt); hydroxychloroquine sulphate (2-{[4-[(7-chloro-4-quinolyl) amino]pentyl]ethylamino}ethanol sulfate (1:1); penicillamine (3-mercapto-D-valine); flurbiprofen ([l,l-biphenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+-.)); cetodolac (1-8 -diethyl- 13, 4,9, tetra hydropyrano-[3-4-13]indole-l-acetic acid; mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydramine hydrochloride (2-diphenyl methoxy-N, N-di-methylethamine hydrochloride).