The United States of America, as Represented by the Secretary, Department of Health and Human Services

Yoram Reiter

Ira H Pastan

David J FitzGerald

This invention provides protease-activatable Pseudomonas exotoxin A-like (&ldquo;PE-like&rdquo;) proproteins. The proproteins comprise (1) a cell recognition domain of between 10 and 1500 amino acids that binds to a cell surface receptor; (2) a modified PE translocation domain comprising an amino acid sequence sufficiently homologous to domain II of PE to effect translocation to a cell cytosol upon proteolytic cleavage, wherein the translocation domain comprises a cysteine-cysteine loop that comprises a protease activatable sequence cleavable by a protease and wherein the cysteine-cysteine loop is substantially un-activatable by furin; (3) optionally, a PE Ib-like domain comprising an amino acid sequence up to 1500 amino acids; (4) a cytotoxicity domain comprising an amino acid sequence substantially homologous to domain III of PE, the cytotoxicity domain having ADP-ribosylating activity; and (5) an endoplasmic reticulum (&ldquo;ER&rdquo;) retention sequence. The invention also provides methods of using these proproteins for killing target cells.

Protease-activatable pseudomonas exotoxin A-like proproteins

This invention provides protease-activatable Pseudomonas exotoxin A-like (&ldquo;PE-like&rdquo;) proproteins. The proproteins comprise (1) a cell recognition domain of between 10 and 1500 amino acids that binds to a cell surface receptor; (2) a modified PE translocation domain comprising an amino acid sequence sufficiently homologous to domain II of PE to effect translocation to a cell cytosol upon proteolytic cleavage, wherein the translocation domain comprises a cysteine-cysteine loop that comprises a protease activatable sequence cleavable by a protease and wherein the cysteine-cysteine loop is substantially un-activatable by furin; (3) optionally, a PE Ib-like domain comprising an amino acid sequence up to 1500 amino acids; (4) a cytotoxicity domain comprising an amino acid sequence substantially homologous to domain m of PE, the cytotoxicity domain having ADP-ribosylating activity; and (5) an endoplasmic reticulum (&ldquo;ER&rdquo;) retention sequence. The invention also provides methods of using these proproteins for killing target cells.

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Yoram Reiter