Tumor necrosis factor superfamily and tnf-like ligand muteins and methods of preparing and using the same

Abstract

The invention features homo-multimers, e.g., homo-trimers, of TNFSF or TNF-like ligand muteins in which each TNFSF ligand or TNF-like ligand mutein monomer contains at least one cysteine residue substitution or insertion that promotes the formation of a disulfide bond with a cysteine residue on a neighboring TNFSF or TNF-like ligand mutein monomer. The invention features methods of producing such TNFSF and TNF-like ligand muteins, pharmaceutical compositions containing such muteins, and methods of using such muteins in cancer immunotherapy, in treating autoimmune and neurological diseases, and in reducing or eliminating the complications and risks of rejection in organ transplantation or tissue or organ repair or regeneration.

1 . A soluble polypeptide comprising all or a portion of a tumor necrosis factor superfamily (TNFSF) or TNF-like ligand, wherein said soluble polypeptide comprises at least one substitution of a surface-exposed, exterior-facing amino acid residue of said TNFSF or TNF-like ligand with a cysteine residue or at least one insertion of a cysteine residue within a region of said TNFSF or TNF-like ligand comprising a surface-exposed, exterior-facing amino acid residue. 
     
     
         2 . A nucleic acid molecule encoding the soluble polypeptide of  claim 1 . 
     
     
         3 . A vector comprising the nucleic acid molecule of  claim 2 . 
     
     
         4 . A host cell comprising the soluble polypeptide of  claim 1  or a nucleic acid molecule encoding the soluble polypeptide. 
     
     
         5 . A pharmaceutical composition comprising a therapeutically effective amount of the soluble polypeptide of  claim 1 , a nucleic acid molecule encoding the soluble polypeptide, a vector comprising the nucleic acid molecule, or a host cell comprising the nucleic acid molecule or the vector. 
     
     
         6 . A method of medical therapy comprising administering the pharmaceutical composition of  claim 5  to a subject in need thereof. 
     
     
         7 . The method of  claim 6 , wherein the subject has an autoimmune disease, a cancer, a neurological disease, an infectious disease, a metabolic disease, a macular disease, muscular atrophy, a disease related to miscarriage, a vascular disease, a disease related to bone loss, an allergy, a blood disorder, a musculoskeletal disorder, a disease related to growth receptors, or obesity, or is in need of tissue or organ repair or regeneration.