Modified ACE2 polypeptides
The present invention relates to modified angiotensin converting enzyme 2 (ACE2) polypeptides and pharmaceutical and analytical uses thereof. In particular, the present invention relates to Zn 2+ depleted-, Zn 2+ free-, mixed metal- and metal ion substituted-ACE2 as well as methods for the manufacture of these variants and uses thereof, such as therapeutic and analytic uses of these ACE2 variants.
1. A pharmaceutical composition comprising a population of angiotensin converting enzyme 2 (ACE2) polypeptides having depleted levels of Zn 2+ in the active sites and a pharmaceutically acceptable carrier or excipient.
2. The pharmaceutical composition according to claim 1 , wherein the ratio of Zn 2+ :ACE2 polypeptides is less than 1:1, preferably ≦0.99:1, ≦0.98:1, ≦0.95:1, ≦0.92:1 or ≦0.90:1.
3. The pharmaceutical composition according to claim 2 , wherein the ratio of Zn 2+ :ACE2 polypeptides is ≦0.50:1, ≦0.45:1, ≦0.40:1, ≦0.35:1, ≦0.30:1, ≦0.25:1, ≦0.20:1, ≦0.15:1, ≦0.10:1, ≦0.05:1, ≦0.02:1, or ≦0.01:1.
4. The pharmaceutical composition according to claim 1 , wherein the ACE2 polypeptides are human ACE2 polypeptides.
5. The pharmaceutical composition according to claim 4 , wherein the amino acid sequences of the ACE2 polypeptides consist of amino acids 1 to 740 of SEQ ID NO: 1, or amino acids 18 to 740 of SEQ ID No: 1, or amino acids 18 to 615 of SEQ ID NO: 1.
6. The pharmaceutical composition according to claim 4 , wherein the ACE2 polypeptides are present in dimeric form.
7. The pharmaceutical composition according to claim 1 , wherein the active sites have no detectable levels of any metal ion.
8. The pharmaceutical composition according to claim 1 , wherein the composition comprises a molar ratio of total zinc or other metal ion to ACE2 of less than 1:1, ≦0.95:1, ≦0.50:1, ≦0.45:1, ≦0.40:1, ≦0.35:1, ≦0.30:1, ≦0.25:1, ≦0.20:1, ≦0.15:1, ≦0.10:1, ≦0.05:1, ≦0.02:1, or ≦0.01:1.
9. The pharmaceutical composition according to claim 1 , wherein the composition comprises metal chelators in a molar excess of 10 to 1000 as compared to the molar amount of zinc in the composition.
10. The pharmaceutical composition as defined in claim 1 for use in treating and preventing a human disease.
11. The pharmaceutical composition as defined in claim 1 for use in the treatment or prevention of a disease selected from hypertension (including high blood pressure), congestive heart failure, chronic heart failure, acute heart failure, contractile heart failure, myocardial infarction, arteriosclerosis, kidney failure, renal failure and lung disease, such as Adult Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI), chronic obstructive pulmonary disease (COPD), pulmonary hypertension, renal fibrosis, chronic renal failure, acute renal failure, chronic renal failure, acute kidney injury and multi-organ dysfunction syndrome.