Claims for Patent: 5,665,593

Summary for Patent: 5,665,593

Title:	Antisense oligonucleotides which combat aberrant splicing and methods of using the same
Abstract:	A method of combatting aberrant splicing in a pre-mRNA molecule containing a mutation is disclosed. When present in the pre-mRNA, the mutation causes the pre-mRNA to splice incorrectly and produce an aberrant mRNA or mRNA fragment different from the mRNA ordinarily encoded by the pre-mRNA. The method comprises hybridizing an antisense oligonucleotide to the pre-mRNA molecule to create a duplex molecule under conditions which permit splicing. The antisense oligonucleotide is one which does not activate RNase H, and is selected to block a member of the aberrant set of splice elements created by the mutation so that the native intron is removed by splicing and the first mRNA molecule encoding a native protein is produced. Oligonucleotides useful for carrying out the method are also disclosed.
Inventor(s):	Kole; Ryszard (Orange County, NC), Dominski; Zbigniew (Orange County, NC)
Assignee:	University of North Carolina (Chapel Hill, NC)
Application Number:	08/379,079
Patent Claims:	1. A method of upregulating expression of a native protein in a cell in vitro, said cell containing a DNA encoding said native protein, which DNA contains a mutation which causes downregulation of said native protein by aberrant splicing in a pre-mRNA, wherein said DNA encodes said pre-mRNA; wherein said pre-mRNA contains a native intron having a first set of splice elements, which native intron is removed by splicing when said mutation is absent to produce a first mRNA encoding said native protein; and wherein said pre-mRNA further contains an aberrant intron different from said native intron having a second set of splice elements, which aberrant intron is removed by splicing when said mutation is present to produce an aberrant second mRNA different from said first mRNA; said method comprising: administering to said cell an antisense oligonucleotide which hybridizes to said pre-mRNA in the nucleus of said cell to create a duplex thereof under conditions which permit splicing, wherein said antisense oligonucleotide does not activate RNase H; and wherein said antisense oligonucleotide blocks a member of said aberrant second set of splice elements; so that said native intron is removed by splicing and said native protein is produced. 2. A method according to claim 1, wherein said cell is an eukaryotic cell selected from the group consisting of yeast, insect, and mammalian cells. 3. A method according to claim 1, wherein said antisense oligonucleotide blocks a mutated splice element. 4. A method according to claim 1, wherein said antisense oligonucleotide blocks a native splice element. 5. A method according to claim 1, wherein said antisense oligonucleotide blocks a cryptic splice element. 6. A method according to claim 1, wherein said antisense oligonucleotide blocks a 5' splice site. 7. A method according to claim 1, wherein said antisense oligonucleotide blocks a 3' splice site. 8. A method according to claim 1, wherein said antisense oligonucleotide blocks a branch point. 9. A method according to claim 1, wherein said native protein is .beta.-globin. 10. A method according to claim 1, wherein said native protein is .beta.-globin. 11. A method according to claim 1, wherein said antisense oligonucleotide is from 8 to 50 nucleotides in length. 12. A method according to claim 1, wherein said antisense oligonucleotide contains a modified internucleotide bridging phosphate residue selected from the group consisting of methyl phosphonates, methyl phosphonothioates, phosphoromorpholidates, phosphoropiperazidates, and phosphoramidates. 13. A method according to claim 1, wherein said antisense oligonucleotide contains a nucleotide having a C1 to C4 lower alkyl substituent at the 2' position thereof.

Details for Patent 5,665,593

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2014-09-09
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2014-09-09
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2014-09-09