Application US20200405829
Massachusetts Institute Of Technology; Wisconsin Alumni Research Foundation
Circular Strained Zymogen Compositions And Methods Of Use
Disclosed herein are circular zymogens of RNase 1 that have a proteolytic cleavage site and are activated by a specific protease. These circular zymogens are useful for treatment of disorders that are characterized by a specific protease (e.g., HIV-1 protease).
Claim CLM-00001. 1. A circular zymogen of ribonuclease 1 (RNase 1) having (i) a truncated C-terminus and (ii) either a wild-type N-terminus or a truncated N-terminus that are connected by a bridge that comprises a proteolytic cleavage site,
wherein the truncated C-terminus has at least 4 residues truncated and the truncated N-terminus has 1 to 6 residues truncated, and/or wherein the circular zymogen is inactivated greater than 104-fold relative to the catalytic activity of wild-type RNase 1, and/or wherein the circular zymogen of RNase 1 is activated by cleavage of the proteolytic cleavage site by a specific protease resulting in a catalytic activity less than 2-fold less than the catalytic activity of wild type RNase 1.
Claim CLM-008-9. 8-9. (canceled)
Claim CLM-00010. 10. A circular zymogen of ribonuclease 1 (RNase 1) having
(a) native termini that are connected by a bridge that comprises a proteolytic cleavage site and has less than 12 residues, and/or (b) a His119 residue, wherein an alpha carbon on the His119 residue is displaced by greater than 1 Å relative to His119 in wild-type RNase 1, and/or (c) a His12 residue, wherein an alpha carbon on the His12 residue is displaced by greater than 1 Å relative to His12 in wild-type RNase 1.
Claim CLM-13-14. 13-14. (canceled)
Claim CLM-00015. 15. A circular zymogen of ribonuclease 1 (RNase 1) comprising an amino acid sequence of His12-X-P-X′-His119, X is a sequence of 1 or more amino acid residues, X′ is a sequence of 1 or more amino acid residues, P is a proteolytic cleavage site, His119 is a histidine-119 residue, and
