Compositions of engineered human arginases and methods for treating cancer
Compositions and methods for the treatment of cancer are described, and, more preferably, to the treatment of cancers that do not express, or are otherwise deficient in, argininosuccinate synthetase, with enzymes that deplete L-Arginine in serum. In one embodiment, the present invention contemplates an arginase protein, such as a human Arginase I protein, comprising at least one amino acid substitution and a metal cofactor, said protein comprising an increased catalytic activity when compared with a native human Arginase I.
1. A composition comprising an isolated human Arginase I and a non-native metal cofactor, wherein the non-native metal cofactor is cobalt and the protein displays a kcat/Km for the hydrolysis of arginine between 400 mM −1 s −1 and 4,000 mM −1 s −1 at pH 7.4, further wherein the human Arginase I has an amino acid sequence as encoded by consisting of SEQ ID NO: 1 NO:13, with or without an N-terminal methionine, the composition being at physiological pH.
2. The composition of claim 1 , wherein the amino acid sequence lacks an N-terminal methionine.
3. The composition of claim 1 , wherein the human Arginase I is covalently linked to polyethylene glycol.
4. A pharmaceutical formulation comprising the composition of claim 1 and a pharmaceutically acceptable excipient and at physiological pH.
5. The formulation of claim 4 , wherein the human Arginase I is covalently linked to polyethylene glycol.
6. A composition comprising an isolated human Arginase I and a non-native metal cofactor, wherein the non-native metal cofactor is cobalt and the protein displays a kcat/Km for the hydrolysis of arginine between 400 mM −1 s −1 and 4,000 mM −1 s −1 at pH 7.4, further wherein the human Arginase I has an amino acid sequence consisting of SEQ ID NO:13, with or without an N-terminal methionine, and is covalently linked to polyethylene glycol, the composition being at physiological pH.
7. A pharmaceutical formulation comprising the composition of claim 6 and a pharmaceutically acceptable excipient and at physiological pH.
8. The composition of claim 6, wherein the PEG is PEG-5000.
9. The composition of claim 6, wherein the arginase is not glycosylated.
10. The formulation of claim 7, wherein the PEG is PEG-5000.
11. The formulation of claim 7, wherein the arginase is not glycoslyated.