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Antisense antiviral agent and method for treating ssRNA viral infection

An RNA virus, virus technology, applied in the field of monitoring the binding of antisense oligomers to viral genome target sites

Inactive Publication Date: 2005-04-06
AVI BIOPHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the absence of reliable cell culture models and appropriate animal models, the development of antisense antiviral agents is quite risky

Method used

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  • Antisense antiviral agent and method for treating ssRNA viral infection
  • Antisense antiviral agent and method for treating ssRNA viral infection
  • Antisense antiviral agent and method for treating ssRNA viral infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0213] Example 1: Antisense inhibits picornavirus (human rhinovirus) in vitro

[0214] The inhibitory effect on rhinovirus 16 of a diaminophosphomorpholino oligomer (PMO) corresponding to the sequence of rhinovirus translation initiation region 14 was used for evaluation. Phosphodiamidate morpholino oligomers (PMO) were synthesized by AVIBioPharma (Corvallis, Oreg.) as described by Summerton and Weller, 1997. Full-length oligomers were more than 90% pure as determined by reversed-phase high pressure liquid chromatography and MALDI TOF mass spectrometry. Lyophilized PMOs were dissolved in sterile 0.9% NaCl solution and filtered through a 0.2 μm Acrodisc filter (Gelman Sciences, Ann Arbor, Mich.) before use in cell culture.

[0215] The PMO contains the nucleic acid sequence targeting rhinovirus 14 and has 3 mismatches with respect to the rhinovirus 16 target sequence. The target sequence (GenBank NC001752 618-637; SEQ ID NO: 5), and the target sequence (SEQ ID NO: 18) are as ...

Embodiment 2

[0223] Example 2: Antisense blocks viruses from porcine kidney (PK-15) and African green monkey kidney (Vero) tissue culture: calicivirus vesicle virus PCV Pan-1 strain and SMSV-13 strain

[0224] The antiviral effects of three phosphorodiamidic morpholino oligomers (PMOs) directed against the ORF1 translation initiation region in Caliciviridae Bursavirus strains Pan-1 and SMSV-13 were evaluated. PMOs were scrape-loaded into two host cells, porcine kidney cells (PK-15) (ATCC No. CCL33) and Vero cells (ATCC No. CCL81 ) (Vero), as described above. The host cells were then exposed to the vesicoviruses Pan-1 and SMSV-13. The measurement and culture of viral load were carried out according to the steps of Example 1.

[0225] For these three PMOs, each included a sequence complementary to the ORF1 initiation region of the Pan-1 sequence (GenBank Accession No. AF091736). The concentration of PMOs in medium without serum is 20.μm. A salt blank and scrambled PMO were used as control...

Embodiment 3

[0232] Example 3: Effect of PMO Antisenses on Feline Calicivirus

[0233] Feline calicivirus becomes feline hemorrhagic virus. Viruses are isolated and propagated on cell culture. Cell cultures were exposed to the virus, followed by detection as described in Example 1. The antisense PMO had the following target sequence: CAG AGT TTG AGA CAT TGT CTC (SEQ ID NO: 32). A one-log reduction in virus titer was observed in cell culture.

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Abstract

The invention provides antisense antiviral compounds and methods of their use in inhibition of growth of viruses of the picornavirus, calicivirus, togavirus, coronavirus, and flavivirus families, as in treatment of a viral infection. In particular embodiments, the virus is an RNA virus from the coronavirus family or a West Nile, Yellow Fever or Dengue virus from the flavivirus family. The antisense antiviral compounds are substantially uncharged oligomers having a targeting base sequence that is substantially complementary to a viral target sequence which spans the AUG start site of the first open reading frame of the viral genome.

Description

field of invention [0001] The present invention relates to an antisense oligomer for treating picornavirus, calicivirus, togavirus or flavivirus infection, and a method for antiviral therapy using the oligomer , and a method of monitoring the binding of an antisense oligomer to a viral genome target site. [0002] references [0003] Agrawal, S. et al., Proc. Natl. Acad. Sci. USA 87(4): 1401-5 (1990). [0004] Alt, M. et al., Hepatology 22(3):707-17(1995). [0005] Alvarez-Salas, L. et al., Antisense Nucleic Acid Drug Dev. 9: 441-450 (1999). [0006] Aurelian, L. et al., Antisense Nucleic Acid Drug Dev. 10: 77-85 (2000). [0007] Bloomers, M., Nuc Acids Res, 22(20): 4187 (1994). [0008] Bonham, M.A. et al., Nucleic Acids Res. 23(7): 1197-1203 (1995). [0009] Boudvillain, M. et al., Biochemistry 36(10):2925-31(1997). [0010] Clarke, G. E., and Lambden, P., J. Infect. Dis. 181, S309-316 (2000). [0011] Cottral, G.E., in: Manual of Standard Methods for Veterinary Micr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53A61K31/7105A61K31/712A61K31/7125A61K38/00A61K48/00A61P31/14C12N15/09C12N15/113C12Q1/68G01N33/566G01N33/569
CPCC12N2310/3145A61K38/00A61K31/712C12N2310/3233C12N15/1131A61K31/7125A61P31/12A61P31/14A61P31/16Y02A50/30
Inventor D·A·斯坦D·E·斯基林P·L·艾弗森A·W·史密斯
Owner AVI BIOPHARMA
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