Tumor-selective e1a and e1b mutants

Patent metadata

Field	Value
Publication number	US-2016017294-A1
Application number	US-201514722021-A
Country	US
Kind code	A1
Filing date	May 26, 2015
Priority date	Mar 2, 2009
Publication date	Jan 21, 2016
Grant date	—

Abstract

Modified E1a regulatory sequences are provided, wherein at least one Pea3 binding site, or a functional portion thereof, is deleted. Also provided are modified E1a sequences that selectively express particular isoforms. Also provided is an E1b-19K clone insertion site. These modified sequences can be used individually, or in combination with one another, to provide tumor-selective expression of proteins.

First claim

1 .- 15 . (canceled)

16 . A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a recombinant adenovirus comprising a modified E1a regulatory sequence, wherein at least one Pea3 binding site, or a functional portion thereof, is deleted.

17 . The pharmaceutical composition of claim 16 , wherein at least one nucleotide in the range of −305 to −141 is retained.

18 . The pharmaceutical composition of claim 16 , wherein at least one of Pea3 II, Pea3 III, Pea3 IV, and Pea3 V, or a functional portion thereof is deleted.

19 . The pharmaceutical composition of claim 18 , wherein at least one of Pea3 II and Pea3 III, or a functional portion thereof is deleted.

20 . The pharmaceutical composition of claim 18 , wherein Pea3 II or a functional portion thereof, and Pea3 III or a functional portion thereof is deleted.

21 . The pharmaceutical composition of claim 18 , wherein at least one of Pea3 IV and Pea3 V, or a functional portion thereof is deleted.

22 . The pharmaceutical composition of claim 16 , wherein said recombinant adenovirus comprises deletion of nucleotides located upstream of a E1a initiation site, wherein the deletion comprises nucleotide −393 to −304, nucleotide −305 to −255, nucleotide −270 to −240, nucleotide −299 to −293, nucleotide −270 to −265, nucleotide −299 to −293, or nucleotide −270 to −265.

23 . The pharmaceutical composition of claim 16 , wherein said recombinant virus selectively expresses an E1a isoform, wherein the sequence encoding the E1a isoform is operably linked to said modified E1a regulatory sequence.

24 . The pharmaceutical composition of claim 23 , wherein said recombinant adenovirus selectively expresses E1a-12S or E1a13S.

25 . The pharmaceutical composition of claim 16 , wherein said recombinant adenovirus substantially excludes expression of an E1a isoform, wherein said sequence encoding said E1a isoform is operably linked to said modified E1a regulatory sequence.

26 . The pharmaceutical composition of claim 25 , wherein said excluded E1a isoform is E1a-12S or E1a-13S.

27 . The pharmaceutical composition of claim 16 , wherein said recombinant adenovirus further comprises a DNA sequence inserted into an E1b-19K insertion site.

28 . The pharmaceutical composition of claim 27 , wherein said insertion site is located between the start site of E1b-19K and the start site of E1b 55K.

29 . The pharmaceutical composition of claim 28 , wherein said E1b-19K insertion site comprises a deletion of 202 base pairs following the start site of E1b-19K.

30 . The pharmaceutical composition of claim 28 , wherein said DNA sequence is a sequence encoding a transgene, a cancer gene, or a mutated DNA sequence.

31 . The pharmaceutical composition of claim 30 , wherein said transgene is a tumor suppressor gene or a functional portion thereof, a toxin gene or a functional portion thereof, a cytokine gene or a functional portion thereof, a pro-drug activating gene or a functional portion thereof, or an apoptotic gene or a functional portion thereof.

32 . The pharmaceutical composition of claim 31 , wherein said transgene is IL-7, IL-12, IL-15, GMCSF, CD40L, CD70, GITRL, OX-40L or a functional portion thereof.

33 . The pharmaceutical composition of claim 31 , wherein said transgene is IL-2, IL-4, IL-5, IL-24, IL-27, CD80, CD137L or a functional portion thereof.

34 . The pharmaceutical composition of claim 30 , wherein said transgene is a mutated p53 sequence.

35 . The pharmaceutical composition of claim 30 , wherein said transgene is tumor necrosis factor, kras or a functional portion thereof.

36 . A method of treating cancer, said method comprising administering to a subject in need thereof a therapeutically effective amount of said pharmaceutical composition of claim 16 or 27 .

37 . A recombinant adenovirus comprising a DNA sequence inserted into an E1b-19K insertion site, wherein said insertion site is located between the start site of E1b-19K and the start site of E1b 55K and wherein said E1b-19K insertion site comprises a deletion of at least about 100 base pairs.

38 . The recombinant adenovirus of claim 37 , wherein said insertion site comprises a deletion of 202 base pairs following said start site of E1b-19K.

39 . The recombinant adenovirus of claim 37 , wherein said DNA sequence is encoding a transgene, a cancer gene, or a mutated DNA sequence.

40 . The recombinant adenovirus of claim 39 , wherein said transgene is a tumor suppressor gene or a functional portion thereof, a toxin gene or a functional portion thereof, a cytokine gene or a functional portion thereof, a pro-drug activating gene or a functional portion thereof, or an apoptotic gene or a functional portion thereof.

41 . The recombinant adenovirus of claim 40 , wherein said transgene is IL-7, IL-12, IL-15, GMCSF, CD40L, CD70, GITRL, OX-40L or a functional portion thereof.

42 . The recombinant adenovirus of claim 40 , wherein said transgene is IL-2, IL-4, IL-5, IL-24, IL-27, CD80, CD137L or a functional portion thereof.

43 . The recombinant adenovirus of claim 39 , wherein said transgene is tumor necrosis factor, kras, or a functional portion thereof.

44 . The recombinant adenovirus of claim 39 , wherein said transgene is a mutated p53 sequence.

45 . A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a recombinant virus of claim 37 .

Assignees

• Univ California

Inventors

• Reid Tony
• Hedjran Farah
• Kumar Shantanu

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C12N2710/10322

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• A61K48/005

  characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered · CPC title

• C12N2710/10343

  viral genome or elements thereof as genetic vector · CPC title

• C12N2830/00

  Vector systems having a special element relevant for transcription · CPC title