Modified oncolytic adenoviruses

A modified replicating adenovirus (Ad) of serotype 5 (Ad5) having lytic activity in target cancer cells comprising:

• a) a E1A gene deletion wherein the deletion is of nucleotides encoding amino acids 923-946 with respect to the wild-type Ad5 sequence;
• b) a 5/3 chimeric substitution of a knob of an adenoviral fiber protein wherein the knob of Ad5 is replaced by the knob of a serotype 3 Ad (Ad3);
• c) a 14.7k gene deletion wherein the deletion is of base pairs 30448-30834 with respect to the wild type Ad5 sequence and wherein the sequence GGA GGA GAT GAC TGA (SEQ ID NO:
• 1) is substituted for GGA GGA GAC GAC TGA (SEQ ID NO:
• 2);
• d) a gp19k gene deletion and a 7.1k gene deletion wherein the deletions are of base pairs 28541-29211 with respect to the wild type Ad5 sequence;
• and
• e) an insertion of a molecule encoding OX40L and a molecule encoding CD40L in the E3B region, replacing the 14.7K gene deletion, wherein the molecule encoding CD40L is inserted immediately downstream from the molecule encoding OX40L using a 2A processing site, and the 2A processing site is preceded by a cleavage site and a SGSG-linker (SEQ ID NO:
• 28) to ensure effective cleavage of OX40L and CD40L.