Compositions and methods of developing oligonucleotides and oligonucleotide analogs having antiviral activity
AssigneeThe Penn State Research Foundation
InventorsJin-Feng Wang, Weihua Pan
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Abstract
Methods of identifying and preparing anti-infectious agent compounds comprising the steps of incubating single stranded nucleic acids or nucleic acid analogs in the presence of infectious agents, selecting for nucleic acids or nucleic acid analogs which bind infectious agents, amplifying selected nucleic acids or nucleic acid analogs, repeating the selection steps and assaying for anti-infection activity are disclosed. Compositions that have anti-infectious activity against Rous sarcoma virus are also disclosed. Methods of identifying anti-tumor agent compounds are also disclosed.
Claims34
A method of identifying a compound which inhibits infection by infectious agents in vitro comprising the steps of
- A) incubating a library of single stranded nucleic acids or nucleic acid analogs in The presence of at least one population of intact infectious agents to effect binding of nucleic acid or nucleic acid analog in said library to said infectious agent, wherein said infectious agent is selected from the group consisting of virus, bacterium, fungus including yeast, and parasite;
- B) separating infectious agent-bound nucleic acid or nucleic acid analog from unbound nucleic acid or nucleic acid analog to obtain a first pool of infectious agent-binding nucleic acid or nucleic acid analog;
- C) amplifying said first infectious agent-binding nucleic acid or nucleic acid analog;
- D) incubating said amplified first infectious agent-binding nucleic acid or nucleic acid analog in the presence of at least one population of intact infectious agents to effect binding of said nucleic acid or nucleic acid analog to said infectious agent;
- E) separating amplified infectious agent-bound nucleic acid or nucleic acid analog from amplified unbound nucleic acid or nucleic acid analog to obtain a second infectious agent-binding nucleic acid or nucleic acid analog;
- F) repeating said amplification step C, said incubation step D and said separation step E from zero up to about thirty times sequentially on each subsequently obtained infectious agent-binding nucleic acid or nucleic acid analog or repeating said steps C through E sufficient times to generate a pool of nucleic acids or nucleic acid analogs which bind to said infectious agent with high affinity;
- and
- G) assaying said subsequently obtained infectious agent-binding nucleic acid or nucleic acid analog for the ability to inhibit infection by said infectious agents in vitro.
A method of identifying a compound which inhibits viral infection in vitro comprising the steps of
- A) incubating a library of RNA and RNA analog molecules in the presence of at least one population of intact viruses to effect binding of said RNA or RNA analog to said viruses;
- B) separating virus-bound RNA or RNA analog from unbound RNA or RNA analog to obtain a first virus-binding RNA or RNA analog;
- C) amplifying said first virus-binding RNA or RNA analog;
- D) incubating said amplified first virus-binding RNA or RNA analog in the presence of at least one population of intact viruses to effect binding of said RNA or RNA analog to said viruses;
- E) separating amplified virus-bound RNA or RNA analog from amplified unbound RNA or RNA analog to obtain a second virus-binding RNA or RNA analog;
- F) repeating said amplification step C, said incubation step D and said separation step E from zero up to about thirty times sequentially on each subsequently obtained virus-binding RNA or RNA analog;
- and
- G) assaying said subsequently obtained virus binding RNA or RNA analog for the ability to bind to said virus an inhibit replication of said virus in vitro.
A method of preparing a compound which inhibits viral infection in vitro comprising the steps of
- A) incubating a library of RNA or RNA flog molecules in the presence of at least one population of intact viruses to effect binding of said RNA or RNA analog to said viruses;
- B) separating virus-bound RNA or RNA analog from unbound RNA or RNA analog to obtain a first virus-bound RNA or RNA analog;
- C) amplifying said first virus-bound RNA or RNA analog;
- D) incubating said amplified first virus-binding RNA or RNA analog in the presence of at least one population of intact viruses to effect binding of said RNA or RNA analog to said viruses;
- E) separating amplified virus-bound RNA or RNA analog from amplified unbound RNA or RNA analog to obtain a second virus-binding RNA;
- F) repeating said amplification step C, said incubation step D and said separation step E from zero up to about thirty times sequentially or repeating said steps C through E sufficient times to generate a pool of nucleic acids or nucleic acid analogs which bind to said infectious agent with high affinity;
- G) assaying said amplified subsequently obtained virus-binding RNA or RNA analog for the ability to bind to said virus and to inhibit replication of said virus in vitro;
- and
- H) isolating and characterizing said subsequently obtained virus-binding RNA or RNA.
An antiRSV composition comprising RNA selected from the group consisting of Sequence B, E, F, G and H (SEQ ID NOS:
- 2, 3, 4, 5 and 6).
Dependencies are inferred from the claim text.