Administration of kynurenine depleting enzymes for tumor therapy

Abstract

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.

What is claimed is: 
     
         1 . A nucleic acid comprising a nucleotide sequence encoding a modified kynureninase enzyme, said modified enzyme having at least one substitution relative to native human kynureninase (see SEQ ID NO: 8), said at least one substitution is at a position selected from the group consisting of: (a) A99, F306, and A436; (b) A99, G112, F306, L337, I405, S   408 ;  (c) G112, F306, L337, and I405; (d) A99, T138, F306, and A436; (e) A99, G112, F306, V339, I405, and 5408; (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, I405, 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; and (j′) F306. 
     
     
         2 . The nucleic acid of  claim 1 , wherein said at least one substitution selected from the group consisting of: (a) A99S, F306L, and A436T; (b) A99V, G112A, F306Y, L337V, I405L, S408N; (c) G112A, F306Y, L337V, and I405L; (d) A99S, T1385, F306L, and A436T; (e) A99V, G112A, F306Y, V339A, I405L, and S408N; (f) A99S and F306L; (g) F306I, L337V, V339I, I405F, and S408T; (h) G112A, F306Y, V339M, and I405L; (i) G1125, F306L, V339T, S408T; (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 I405L; (p) A99I, G112A, F306Y, I405L, S408N, 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; and (j′) F306Y. 
     
     
         3 . The nucleic acid of  claim 1 , wherein the encoded modified kynureninase further comprises a heterologous peptide segment. 
     
     
         4 . The nucleic acid of  claim 1 , wherein the nucleic acid is codon optimized for expression in bacteria, fungus, insects, or mammals. 
     
     
         5 . An expression vector comprising the nucleic acid of  claim 1 . 
     
     
         6 . A host cell comprising the nucleic acid of  claim 1 . 
     
     
         7 . The host cell of  claim 6 , wherein the host cell is a bacterial cell, a fungal cell, an insect cell, or a mammalian cell. 
     
     
         8 . A pharmaceutical formulation comprising the nucleic acid of  claim 1  in a pharmaceutically acceptable carrier. 
     
     
         9 . The formulation of  claim 8 , wherein the nucleic acid encoding the modified kynureninase is codon optimized for expression in bacteria, fungus, insects, or mammals. 
     
     
         10 . The formulation of  claim 8 , wherein the nucleic acid is in an expression vector. 
     
     
         11 . A transgenic T cell comprising an expressed chimeric antigen T-cell receptor (CAR) and an expressed modified kynureninase enzyme encoded by the nucleic acid of  claim 1 . 
     
     
         12 . The cell of  claim 11 , wherein the cell is a human T cell.