Variants of thymidine kinase, nucleic acids encoding them, and methods of using them
AssigneeAventis Pharma S.A.
InventorsJoël Crouzet, Francis Blanche, Michel Couder, Béatrice Cameron
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Abstract
The present invention relates to a nucleic acid sequence characterized in that it is derived from the wild nucleic acid sequence coding for a thymidine kinase, said nucleic acid sequence having at least one mutation in the region corresponding to the ATP binding site and conveniently a second mutation in the N-terminal region and/or C-terminal region. It also relates to variants of the wild thymidine kinase and their use in genic therapy.
Claims37
A nucleic acid coding for a herpes simplex virus type 1 thymidine kinase variant, wherein the variant has a mutation in the region corresponding to the ATP-binding site, and an additional one or more mutations in the N-terminal or C-terminal regions, the mutations selected from the following group or from a mutation resulting in a degenerate coding variant of any in the following group:
- adenine substituting for the guanine at position 180 (G180A);
- adenine substituting for the guanine at position 16 (G16A);
- adenine substituting for the guanine at position 28 (G28A);
- adenine substituting for the guanine at position 30 (G30A);
- adenine substituting for the guanine at positions 28 and 30 (G28A and G30A);
- thymine substituting for the cytosine at position 591 (C591T);
- thymine substituting for the cytosine at position 892 (C892T);
- thymine substituting for the cytosine at positions 591 and 892 (C591T and C892T);
- adenine substituting for the guanine at position 1010 (G 1010A);
- adenine substituting for the guanine at position 1011 (G1011A);
- adenine substituting for the guanine at positions 1010 and 1011 (G1010A and G1011A);
- and adenine substituting for the guanine at positions 1010 and 1011 and a thymine
- substituting for the cytosine at positions 591 and 892 (G1010A;
- G1011A;
- C591T;
- and
- C892T).
A thymidine kinase variant comprising an ATP-binding site motif of SEQ ID No:
- 1 or SEQ ID No:
- 2, having a substitution mutation in SEQ ID No:
- 1 or SEQ ID No:
- 2, and having an additional one or more substitution mutations in the C-terminal or N-terminal regions, the mutations selected from the following sites corresponding to herpes simplex virus type 1 thymidine kinase:
- methionine at position 60;
- alanine at position 10;
- glycine at position 6;
- arginine at position 337;
- methionine at position 60 and alanine at position 10;
- methionine at position 60 and alanine at position 10 and arginine at position 337;
- methionine at position 60 and glycine at position 6.
A method of using a composition of claim 36, comprising administering the composition in an amount effective to transform a prodrug into a toxic agent to eliminate a dividing cell.
Dependencies are inferred from the claim text.