Arginine deiminase encapsulated inside erythrocytes and their use in treating cancer and arginase-1 deficiency

1. A method of treatment of arginase-1 deficiency, comprising administering to a patient in need thereof an effective amount of a pharmaceutical composition comprising a magnesium-independent arginine deiminase (ADI) encapsulated into erythrocytes and a pharmaceutically acceptable vehicle.

2. The method according to claim 1 , wherein said composition is a suspension having an osmolarity of between 270 and 350 mOsm/L; and wherein the ADI further does not depend upon iron or manganese, or is co-factor-independent.

3. The method according to claim 1 , wherein the pharmaceutically acceptable vehicle is a preservative solution comprising NaCl and Adenine.

4. The method according to claim 1 , wherein the ADI is from Mycoplasma arginini.

5. The method according to claim 1 , wherein the ADI comprises or consists essentially of the amino acid sequence of SEQ ID NO: 1.

6. The method according to claim 1 , wherein the ADI comprises an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO: 1.

7. The method according to claim 1 , wherein the concentration of encapsulated ADI is from 0.1 to 7 mg/ml.

8. The method of claim 1 , wherein the pharmaceutical composition is formulated as a dose having a volume from 10 to 250 ml.

9. The method of claim 8 , wherein the dose comprises from 0.01 mg/kg to 500 mg/kg of encapsulated ADI per kg patient body weight.

10. A method of treatment of arginase- 1 deficiency or arginine-dependent cancers, treating or preventing of septic shock, inhibiting angiogenesis and treating angiogenesis associated diseases, comprising administering to a patient in need thereof an effective amount of a suspension of erythrocytes encapsulating a magnesium-independent arginine deiminase from Mycoplasma arginini.

11. The method of claim 10 , wherein said ADI comprises the amino acid sequence set forth in SEQ ID NO: 1.

12. The method of claim 10 , wherein the ADI further neither requires iron nor manganese, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintain depleted levels of arginine for more than 10 days.

13. The method of claim 12 , wherein the ADI does not require any co-factor, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintains depleted levels of arginine for more than 10 days.

14. The method of claim 1 , further comprising reducing pathological plasma or whole blood arginine levels in a patient or subject suffering from Arginase 1 deficiency to normal physiological or near-normal physiological plasma or whole blood arginine levels, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintain depleted levels of arginine for more than 10 days.

15. The method of claim 1 , wherein the composition reduces pathological levels of arginine by at least about 50, 60, 70 or about 80% for a period of at least about 6, 7, 8, 9, 10 or 11 days post administration of a single dose of the composition; and wherein the ADI is also iron-independent and/or manganese-independent and/or co-factor independent.

16. The method of claim 1 , wherein the composition is characterized by any or all of the following ranges of values across the indicated parameters: hematocrit (%): about 48 to about 51; corpuscular volume (fL): about 82 to about 88; corpuscular hemoglobin (g/dL): about 27 to about 30; RBC count (10 6 /μl): about 5.5 to about 6.2; total hemoglobin (g/dL): about 13.0 to about 14.5; extracellular Hb (g/dL): about 0.1 to about 0.2; intra-erythrocyte concentration of ADI (mg/mL of RBC): about 0.8 to about 1.6; intra-erythrocyte activity of ADI (U/mL): about 42 to about 49 or about 65 to about 74; extracellular activity (%): about 0.0 to about 4.0; intracellular activity (%): about 96 to about 100; and/or entrapment yield of ADI (%): about 27 to about 34, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintains depleted levels of arginine for more than 10 days.

17. The method of claim 2 , further comprising reducing pathological plasma or whole blood arginine levels in a patient or subject suffering from Arginase 1 deficiency to normal physiological or near-normal physiological plasma or whole blood arginine levels, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintains depleted levels of arginine for more than 10 days.

18. The method of claim 2 , wherein the composition reduces pathological levels of arginine by at least about 50, 60, 70 or about 80% for a period of at least about 6, 7, 8, 9, 10 or 11 days post administration of a single dose of the composition; and wherein the ADI is also iron-independent and/or manganese-independent and/or co-factor independent.

19. The method of claim 2 , wherein the composition is characterized by any or all of the following ranges of values across the indicated parameters: hematocrit (%): about 48 to about 51; corpuscular volume (fL): about 82 to about 88; corpuscular hemoglobin (g/dL): about 27 to about 30; RBC count (10 6 /μL): about 5.5 to about 6.2; total hemoglobin (g/dL): about 13.0 to about 14.5; extracellular Hb (g/dL): about 0.1 to about 0.2; intra-erythrocyte concentration of ADI (mg/mL of RBC): about 0.8 to about 1.6; intra-erythrocyte activity of ADI (U/mL): about 42 to about 49 or about 65 to about 74; extracellular activity (%): about 0.0 to about 4.0; intracellular activity (%): about 96 to about 100; and/or entrapment yield of ADI (%): about 27 to about 34, and wherein a single administration of erythrocytes encapsulating the magnesium-independent ADI maintains depleted levels of arginine for more than 10 days.