Truncated guinea pig L-asparaginase variants and methods of use
Variant guinea pig L-asparaginases which are truncated and humanized are described as are fusion proteins containing the L-asparaginase and use of the L-asparaginases in the treatment of cancers such as acute lymphoblastic leukemia and acute myeloid leukemia.
1. A Guinea pig L-Asparaginase (GpA) variant, wherein said GpA variant comprises an amino acid sequence having at least 85% sequence identity to the sequence of amino acid residues 1 to 359 of SEQ ID NO: 1, and has one or more amino acid substitutions relative to SEQ ID NO: 1, said amino acid substitutions being selected from the group consisting of:
H10R, Q23R, K25E, K48E, Q52R, Q54R, P57S, DS8E, H59D, A60T, A62V, D91A, D92E, K98Q, E101K, Q108H, G122A, H1340, R147H, K193R, D217E, N233S, H236Q, S250A, Q288E, R301Q, E344D, L360P, T362S, A363V, D364E, L365E, H366R, Q367R and S368P; and
optionally further having one or more amino acid substitutions relative to SEQ ID NO: 1 selected from the group consisting of C198A, C198S, C198V, E49C, R52C, K225C, Q257C, Q281C, and E340C.
2. The GpA variant of claim 1 , wherein the variant is truncated at a position between amino acid residues corresponding to positions 359 and 396 of SEQ ID NO: 1.
3. The GpA variant of claim 1 , wherein the variant is truncated at the position corresponding to position 369 of SEQ ID NO: 1.
4. The GpA variant of claim 1 , wherein the variant has a catalytic activity equal to or greater than wild-type GpA comprising the amino acid sequence of SEQ ID NO: 1.
5. The GpA variant of claim 1 , further comprising a histidine tag, a SUMO tag, an albumin-binding domain, or a combination thereof.
6. A nucleic acid molecule comprising a nucleotide sequence encoding the GpA variant of claim 1 .
7. An expression vector comprising the nucleic acid molecule of claim 6 .
8. An isolated host cell comprising the nucleic acid molecule of claim 6 .
9. A pharmaceutical composition comprising the GpA variant of claim 1 and a pharmaceutically acceptable excipient.
10. A method of treating cancer in a subject comprising administering an effective amount of the GpA variant of claim 1 to a subject in need thereof, thereby treating the subject's cancer.
11. The method of claim 10 , wherein the cancer is selected from non-Hodgkin's lymphoma, leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, B cell lymphoma, Burkitt's lymphoma, chronic myelocytic leukemia, chronic lymphocytic leukemia, and hairy cell leukemia.
12. A Guinea pig L-Asparaginase (GpA) variant comprising the amino acid sequence of SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, or SEQ ID NO: 86.
13. The GpA variant of claim 12 , further comprising a histidine tag, a SUMO tag, an albumin-binding domain, or a combination thereof.
14. The GpA variant of claim 12 , further comprising three tandem soluble domains of TRAIL.
15. The GpA variant of claim 14 , wherein the soluble domains of TRAIL comprise residues 115-281 of human TRAIL.