Administration of kynurenine depleting enzymes for tumor therapy
Methods and compositions related to the use of a protein with kynureninase activity are described. For example, in certain aspects there may be disclosed a modified kynureninase capable of degrading kynurenine. Furthermore, certain aspects of the invention provide compositions and methods for the treatment of cancer with kynurenine depletion using the disclosed proteins or nucleic acids.
1. An isolated, modified human kynureninase enzyme, said isolated, modified human kynureninase enzyme having at least one substitution relative to native human kynureninase comprising the amino acid sequence of SEQ ID NO: 8, wherein said at least one substitution is at positions: (a) A99, F306, and A436; (b) A99, G112, F306, L337, I405, S408; (c) G112, F306, L337, and I405; (d) A99, T138, F306, and A436; (e) A99, G112, F306, V339, I405, and S408; (f) A99 and F306; (g) F306, L337, V339, I405, and S408; (h) G112, F306, V339, and I405; (i) G112, F306, V339, S408; (k) F71, A99, G112, T138, F306, L337, V339, I405, S408, and A436; (1) A99, G112, F306, L337, V339, I405, and S408; (m) A436; (n) A99, G112, T138, V339, and I405; (p) A99, G112, F306, 1405, S408, and A436; (q) F71, A99, I131, F249, and L322; (r) A99, I131, F249, E259, and F306; (s) F71, A99, and E259; (t) F71, A99, S167, and E259; (u) I131, F249, and S274; (v) L59, G112, F306, V339, I405, and S408; (w) I110 and F306; (x) A99, I131, F249, and E259; (y) F71, E259, and L322; (z) H41, Q175, and A436; (a′) A99, I131, and F249; (b′) I131 and F249; (c′) T138 and A436; (d′) T138; (e′) F71, A99, I131, E259, and V303; (f) A99, G112, F306, V339, I405, and S408; (g′) F71, A99, I131, E259, and A282; (h′) F71, F249, E259, and V303; (i′) I110; or (j′) F306.
2. The enzyme of claim 1 , wherein said at least one substitution is at positions: (a) A99S, F306L, and A436T; (b) A99V, G112A, F306Y, L337V, I405L, 5408N; (c) G112A, F306Y, L337V, and I405L; (d) A99S, T138S, F306L, and A436T; (e) A99V, G112A, F306Y, V339A, I405L, and 5408N; (f) A99S and F306L; (g) F306I, L337V, V339I, I405F, and S408T; (h) G112A, F306Y, V339M, and I405L; (i) G112S, F306L, V339T, 5408T; (j) G112A, F306Y, V339S, I405L; (k) F71L, A99I, G112A, T138S, F306Y, L337V, V339I, I405L, S408N, and A436T; (1) A99V, G112A, F306Y, L337V, V339I, I405F, and S408N; (m) A436T; (n) A99V, G112A, T138S, V339A, and I405F; (o) G112S, F306Y, V339T, and 1405L; (p) A99I, G112A, F306Y, I405L, 5408N, and A436T; (q) F71L, A99I, I131V, F249W, and L322P; (r) A99I, I131V, F249W, E259P, and F306L; (s) F71L, A99I, and E259P; (t) F71L, A99I, S167T, and E259P; (u) I131M, F249W, and S274G; (v) L59M, G112S, F306Y, V339A, I405L, and S408N; (w) I110L and F306L; (x) A99I, I131V, F249W, and E259P; (y) F71L, E259P, and L322P; (z) H41R, Q175L, and A436T; (a′) A99I, I131V, and F249W; (b′) I131V and F249W; (c′) T138S and A436T; (d′) T138S; (e′) F71L, A99I, I131V, E259P, and V303S; (f) A99F, G112A, F306Y, V339A, I405L, and S408N; (g′) F71L, A99I, I131V, E259P, and A282P; (h′) F71L, F249W, E259P, and V303S; (i′) I110L; or (j′) F306Y.
3. The enzyme of claim 1 , further comprising a heterologous peptide segment.
4. The enzyme of claim 1 , wherein the enzyme is coupled to polyethylene glycol (PEG).
5. The enzyme of claim 4 , wherein the enzyme is coupled to the PEG via one or more Lys or Cys residues.
6. A pharmaceutical formulation comprising the isolated, modified kynureninase of claim 1 in a pharmaceutically acceptable carrier.
7. The formulation of claim 6 , wherein the isolated, modified kynureninase further comprises a heterologous peptide segment.
8. The formulation of claim 6 , wherein the isolated, modified kynureninase is coupled to polyethylene glycol (PEG).
9. The formulation of claim 8 , wherein the isolated, modified kynureninase is coupled to PEG via one or more Lys or Cys residues.
10. The enzyme of claim 1 , wherein the isolated, modified kynureninase has at least 90% sequence identity to SEQ ID NO:8.
11. The enzyme of claim 1 , wherein the isolated, modified kynureninase has at least 95% sequence identity to SEQ ID NO:8.
12. The enzyme of claim 1 , wherein the isolated, modified kynureninase further comprises one or more chemical modifications.
13. The enzyme of claim 12 , wherein the one or more chemical modifications is at a substrate recognition site.
14. The enzyme of claim 1 , wherein the isolated, modified kynureninase is conjugated to an antibody.
15. The enzyme of claim 14 , wherein the antibody is a scFv antibody.
16. The enzyme of claim 14 , wherein the antibody is an anti-CTLA4 antibody, anti-PD1 antibody, or anti-PDL1 antibody.