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(12) United States Patent (10) Patent No.: US 7,527,801 B2 Coit Et Al

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(12) United States Patent (10) Patent No.: US 7,527,801 B2 Coit Et Al USOO7527801B2 (12) United States Patent (10) Patent No.: US 7,527,801 B2 Coit et al. (45) Date of Patent: May 5, 2009 (54) NOROVIRUS AND SAPOVIRUS ANTIGENS Genbank Accession # AAQ63158, ORF3 of Norwalk virus, pub lished Aug. 31, 2003.* (75) Inventors: Doris Coit, Petaluma, CA (US); Glass et al., Two Nonoverlapping Domains on the Norwalk Virus Michael Houghton, Danville, CA (US); Open Reading Frame 3 (ORF3) Protein Are Involved in the Forma Colin McCoin, Castro Valley, CA (US); tion of the Phosphorylated 35K Protein and in ORF3-Capsid Protein Interactions, 2003, Journal of Virology, vol. 77. No. 6, pp. 3569 Angelica Medina-Selby, San Francisco, 3577.: CA (US); Michael Vajdy, Orinda, CA Harrington et al., Systemic, Mucosal, and Heterotypic Immune (US) Induction in Mice Inoculated with Venezuelan Equine Encephalitis Replicons ExpressingNorwalkVirus-Like Particles, 2002, Journal of (73) Assignee: Novartis Vaccines and Diagnostics, Virology, vol. 76, No. 2, pp. 730-742.* Inc., Emeryville, CA (US) Asanaka, et al., “Replication And Packaging Of Norwalk Virus RNA In Cultured Mammalian Cells. PNAS USA 102: 10327-10332 (*) Notice: Subject to any disclaimer, the term of this (2005). patent is extended or adjusted under 35 Belliot, et al., “In Vitro Proteolytic Processing Of The MD 145 U.S.C. 154(b) by 0 days. Norovirus ORF1 Nonstructural Polyprotein Yields Stable Precursors And Products Similar To Those Detected In Calicivirus-Infected (21) Appl. No.: 11/603.913 Cells.”J Virol 77: 10957-10974 (2003). Chakravarty, et al., “Evolutionary Trace Residues. In Noroviruses: (22) Filed: Nov. 22, 2006 Importance In Receptor Binding, Antigenicity, Virion Assembly, And Strain Diversity.”J Virol 79:554-568 (2005). (65) Prior Publication Data Chang, et al., “Bile Acids Are Essential For Porcine Enteric Calicivirus Replication. In Association With Down-Regulation Of US 2007/O2O7526A1 Sep. 6, 2007 Signal Transducer And Activator Of Transcription 1.” PNAS USA 101:8733-8738 (2004). Related U.S. Application Data Chen, et al., “Inter- And Intragenus Structural Variations. In (60) Provisional application No. 60/739,217, filed on Nov. Caliciviruses And Their Functional Implications.” J Virol 78:6469 22, 2005. 6479 (2004). Fankhauser, et al., “Molecular Epidemiology Of “Norwalk-Like Viruses' In Outbreaks Of Gastroenteritis In The United States.' J (51) Int. Cl. Infect Dis 178: 1571-1578 (1998). AOIN 63/04 (2006.01) Fankhauser, et al., “Epidemiologic And Molecular Trends Of A6 IK 39/38 (2006.01) “Norwalk-Like Viruses' Associated With Outbreaks Of A6 IK 39/2 (2006.01) Gastroenteritis In The United States.”J Infect Dis 186:1-7 (2002). A 6LX 39/25 (2006.01) Farkas, et al., “Genetic Diversity Among Sapoviruses.” Arch Virol CI2P 2L/04 (2006.01) 149: 1309-1323 (2004). CI2N 5/02 (2006.01) CI2N 15/74 (2006.01) (Continued) A61 K39/187 (2006.01) Primary Examiner Bruce Campell (52) U.S. Cl. .............. 424/216.1; 424/184.1; 424/199.1; Assistant Examiner Benjamin P Blumel 424/204.1; 424/93.51; 435/483; 435/325: (74) Attorney, Agent, or Firm Regina Bautista; Roberta 435/70.1 Robins (58) Field of Classification Search ....................... None See application file for complete search history. (57) ABSTRACT (56) References Cited Immunogenic compositions that elicit immune responses U.S. PATENT DOCUMENTS against Norovirus and Sapovirus antigens are described. In 6, 194,560 B1* 2/2001 Arntzen et al. ............. 536,23.7 particular, the invention relates to polynucleotides encoding 6,551,820 B1* 4/2003 Mason et al. ............ 435/320.1 one or more capsid proteins or other immunogenic viral 2003/0129588 A1 7/2003 Estes et al. polypeptides from one or more strains of Norovirus and/or 2005/O152911 A1 7/2005 Hardy Sapovirus, coexpression of such immunogenic viral polypep tides with adjuvants, and methods of using the polynucle FOREIGN PATENT DOCUMENTS otides in applications including immunization and produc JP 2002O2O399. A 1, 2002 tion of immunogenic viral polypeptides and viral-like WO WO93,03769 A1 3, 1993 particles (VLPs). Methods for producing Norovirus- or WO WO94,05700 A2 3, 1994 Sapovirus-derived multiple epitope fusion antigens or WO WO 2005/030806 A2 4/2005 polyproteins and immunogenic compositions comprising one WO WO 2005/032457 A2 4/2005 or more immunogenic polypeptides, polynucleotides,VLPs, and/or adjuvants are also described. The immunogenic com OTHER PUBLICATIONS positions of the invention may also contain antigens other than Norovirus or Sapovirus antigens, including antigens that Baric et al., Expression and Self-Assembly of Norwalk Virus Capsid Protein from Venezuelan Equine Encephalitis Virus Replicons, 2002, can be used in immunization against pathogens that cause Journal of Virology, vol. 76, No. 6, pp. 3023-3030.* diarrheal diseases, such as antigens derived from rotavirus. Genbank Accession # BAA83413, ORF2 of Norwalk virus, pub lished Dec. 2, 2003.* 7 Claims, 38 Drawing Sheets US 7,527,801 B2 Page 2 OTHER PUBLICATIONS Lindesmith, et al., “Cellular and humoral immunity following Snow Green, et al., “Expression And Self-Assembly Of Recombinant Mountain virus challenge.”J Virol 79:2900-2909 (2005). Capsid Protein From The Antigenically Distinct Hawaii Human Nicollier-Jamot, et al., “Recombinant Virus-Like Particles Of A Calicivirus.” J Clin Microbiol 35:1909-1914 (1997). Norovirus (Genogroup II Strain) Administered Intranasally And Harrington, et al., “Norovirus Capture With Histo-Blood Group Anti Orally With Mucosal Adjuvants LT And LT(R192G) In BALB/C gens Reveals NovelVirus-Ligand Interactions.”.JVirol 78:3035-3045 Mice Induce Specific Humoral And Cellular Th1/Th2-Like Immune (2004). Responses.” Vaccine 22: 1079-1086 (2004). Hutson, et al., “Norwalk Virus-Like Particle Hemagglutination By Oka, et al., “Proteolytic Processing Of Sapovirus ORF1 Polyprotein. Binding To H Histo-Blood Group Antigens,” J Virol 77:405-415 J Virol 79:7283-7290 (2005). (2003). Schuffenecker, et al., “Genetic classification of 'Sapporo-like Jiang, et al., "Sapporo-Like Human Caliciviruses Are Genetically viruses’.” Arch Virol 146(11):2115-2132 (2001). and Antigenically Diverse.” Arch Virol 142: 1813-1827 (1997). Subekti, et al., “Experimental Infection Of Macaca Nemestrina With Jiang, et al., “Sequence And Genomic Organization Of Norwalk A Toronto Norwalk-Like Virus Of Epidemic Viral Gastroenteritis.”J Virus.” Virology 195:51-61 (1993). Medical Virology 66:400-406 (2002). Johnson, et al., “Multiple-Challenge Study Of Host Susceptibility To Taube, et al., “Generation Of Recombinant Norovirus-Like Particles Norwalk Gastroenteritis In US Adults.” J Infect Dis 161:18-21 (VLP) In The Human Endothelial Kidney Cell Line 293T.” Arch Virol (1990). 150: 1425-1431 (2005). Lindesmith, et al., “Human Susceptibility And Resistance To Norwalk Virus Infection.” Nature Medicine 9:548-533 (2003). * cited by examiner U.S. Patent May 5, 2009 Sheet 1 of 38 US 7,527,801 B2 VIºunãIA U.S. Patent May 5, 2009 Sheet 2 of 38 US 7,527,801 B2 {{I?InáH 1.98 988 4,96 9/6 IÇIT 09TT /.9GT. 98GT U.S. Patent May 5, 2009 Sheet 3 of 38 US 7,527,801 B2 OIounãIJ 600Z 8ZOZ 660Z 8ITZ 68TZ 80Z U.S. Patent May 5, 2009 Sheet 7 Of 38 US 7,527,801 B2 GIZ9.InáH ?6*0.9g9G=nuô?aM-TOWp?3eTSUIeTJ, U.S. Patent May 5, 2009 Sheet 9 Of 38 US 7,527,801 B2 0LT09T 06T08T „IzºunãIJ ZIZ U.S. Patent May 5, 2009 Sheet 10 of 38 US 7,527,801 B2 S dç’AN U.S. Patent May 5, 2009 Sheet 11 of 38 US 7,527,801 B2 I#ZJIo’AN’ZL?Sd †9.InáH IBQXIIIpu?H U.S. Patent May 5, 2009 Sheet 12 of 38 US 7,527,801 B2 L]+[] C+ZJuo’AN’Z?dSd LL]+[()] U.S. Patent May 5, 2009 Sheet 13 of 38 US 7,527,801 B2 IIES I#ZJIO'AN'pd 99.InáH U.S. Patent US 7,527,801 B2 9+ZJIO'AN'pd L9.InáH U.S. Patent May 5, 2009 Sheet 16 of 38 US 7,527,801 B2 i U.S. Patent May 5, 2009 Sheet 17 Of 38 US 7,527,801 B2 IIES 0I9.In?š?H ("VAL"V906LEOd |Z#ZJIO’AN U.S. Patent US 7,527,801 B2 w or cN v w cy CN ve O uuOgye 03ue duo SqW U.S. Patent May 5, 2009 Sheet 20 of 38 US 7,527,801 B2 U.S. Patent May 5, 2009 Sheet 21 of 38 US 7,527,801 B2 Figure 14A NV.orf2: modified polynucleotide sequence of orf2 (SEQ ID NO:1) aa.gcttacaa aacaaaatga t gatggCg to taaggacgCt a Cat CaagCg tygatggCgC 60 tag togg.cgct gg toagttgg taccggagglt taatgCttct gaCCCtcttg Caatgg accc 120 tg tag caggt tottcgacag cag togcgac tgctggacaa gttaatcCta ttgatccctg 180 gataatcaat aattttgttgc aag CCCCCCa aggtgaattit actattt coc Caaataatac 240 ccccgg togat gttttgtttg atttgagttt ggg tocccat cittaatcctt tottgcticca 300 totaticacaa atgtataatg gttgggttgg taacatgaga g toaggatta togttggctgg 360 taatgcc titt actg.cgggga agataatag t t to Ctgcata ccc.cctggitt ttgg ttcaca 420 taatct tact a tag cacaag caactct Ctt toca Catgtg attgctgatg t taggactCt 480 agacccCatt gaggtgCCtt tdgaagatgt taggaatgtt Ctct ttcata ataatgatag 540 aaatcaacaa accatgCgCC ttgttgttg Cat gCtgtaCaCC CCCC to CJCa Ctggtggtgg 600 tactggtgat tcttttgtag ttgCagg gCQ agittatgact togCCCCagtC Ctgattittaa 660 tttcttgttt ttag toccitc Ctacgg tyga gCagaaaacc aggCCCttca cacticcolaaa 720 totgccattg agttctotgt Ctaacticacg togCCCCtcto coaatcagta gitatcgg cat 780 ttcc.ccagac aatgtccaga gtgtgcagtt coaaaatggit cqgtg tactic toggatgg.ccg. 840 cctggttggc accaccocag tt to attgtc. acatgttgcc aagataagag ggacctocaa 900 tggcactgta atcaacctta ctgaattgga tiggCacaccC tittcaccctt ttgaggg.ccc 960 tgccoccatt gggtttccag acct cqgtgg ttgttgattgg catattaata toacacagtt 1020 tggccattct agccaga CCC ag tatgatgt agaCaCCaCC CCtgacaCtt ttgtCCCCCa 1080 tcttggttca attcaggcaa atgg Cattgg Cagtgg taat tatgttggtg tt Cittagctg 1140 gattitccc.ca ccatcacacc cg totggctC CCaagttgaC Ctttggaaga toccCaatta 1200 tggg toaagt attacggagg CaacaCatct agCCCCttct gtatacCCCC Ctgg tttcqg 1260 agagg tattg g tottct tca to tccaagat goCagg toct gg togCttata atttgccCtg 1320 totattacca caagagtaca tt toacatct togctagtgaa caag.cccCta Ctgtaggtga 1380 ggctg.ccctg.
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