Therapeutic nuclease compositions and methods

Abstract

Hybrid nuclease molecules and methods for treating an immune-related disease or disorder in a mammal, and a pharmaceutical composition for treating an immune-related disease in a mammal.

1 . A hybrid nuclease molecule comprising a first nuclease domain and a modified Fc domain, wherein the first nuclease domain is operatively coupled to the Fc domain. 
     
     
         2 .- 63 . (canceled) 
     
     
         64 . The hybrid nuclease molecule of  claim 1  wherein the Fc domain is modified to decrease binding to Fcγ receptors or complement proteins or both. 
     
     
         65 . The hybrid nuclease molecule of  claim 64  which has a reduced effector function selected from the group consisting of opsonization, phagocytosis, complement dependent cytotoxicity, and antibody-dependent cellular cytotoxicity. 
     
     
         66 . The hybrid nuclease molecule of  claim 1 , further comprising a second nuclease domain operatively coupled to the Fc domain. 
     
     
         67 . The hybrid nuclease molecule of  claim 1 , wherein the first nuclease domain is operatively coupled to the Fc domain via a first linker domain 
     
     
         68 . The hybrid nuclease molecule of  claim 67 , wherein the first linker domain is a polypeptide linker, such as a gly-ser linker. 
     
     
         69 . The hybrid nuclease molecule of  claim 66 , wherein the second nuclease domain is operatively coupled to the Fc domain via a second linker domain. 
     
     
         70 . The hybrid nuclease molecule of  claim 69 , wherein the second linker domain is a polypeptide linker, such as an NLG peptide. 
     
     
         71 . The hybrid nuclease molecule of  claim 1 , wherein the first nuclease domain comprises an RNase. 
     
     
         72 . The hybrid nuclease molecule of  claim 71 , wherein the RNase is a human RNase, such as a human pancreatic RNase A. 
     
     
         73 . The hybrid nuclease molecule of  claim 1 , wherein the Fc domain comprises a human immunoglobulin Fc domain, such as a human IgG1 Fc domain. 
     
     
         74 . The hybrid nuclease molecule of  claim 73 , wherein the Fc domain comprises a hinge domain, a CH2 domain and a CH3 domain. 
     
     
         75 . The hybrid nuclease molecule of  claim 66 , wherein the second nuclease domain comprises a DNase. 
     
     
         76 . The hybrid nuclease molecule of  claim 75 , wherein the DNase is selected from the group consisting of a Type I human DNase, a human DNase 1L3, or human TREX1. 
     
     
         77 . The hybrid nuclease molecule of  claim 1  comprising a polypeptide, wherein the first nuclease domain comprises an amino acid sequence of a mature peptide of a human RNase pancreatic precursor set forth in SEQ ID NO:149, or an RNase comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:149. 
     
     
         78 . The hybrid nuclease molecule of  claim 1 , wherein the Fc domain is a human IgG1 Fc domain comprising an amino acid sequence set forth in SEQ ID NO:145, or an Fc domain comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:145. 
     
     
         79 . The hybrid nuclease molecule of  claim 66  comprising a polypeptide, wherein the second nuclease domain comprises a human DNase comprising an amino acid sequence set forth in SEQ ID NO:143, or a DNase comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:143. 
     
     
         80 . The hybrid nuclease molecule of  claim 1 , wherein the Fc domain comprises a modified hinge domain comprising at least one substitution. 
     
     
         81 . The hybrid nuclease molecule of  claim 80 , wherein the modified hinge domain comprises at least one cysteine substitution, such as a substitution with serine. 
     
     
         82 . The hybrid nuclease molecule of  claim 1 , wherein the Fc domain comprises a modified CH2 domain comprising at least one substitution. 
     
     
         83 . The hybrid nuclease molecule of  claim 82 , wherein the modified CH2 domain comprises at least one substitution of an amino acid associated with Fcγ receptor binding or complement protein binding or both. 
     
     
         84 . The hybrid nuclease molecule of  claim 83 , wherein the substitution is selected from the group consisting of P238S, P331S, N297S or combination thereof. 
     
     
         85 . A hybrid nuclease molecule comprising a polypeptide, wherein the polypeptide comprises an amino acid sequence of a mature peptide of a human RNase pancreatic precursor operatively linked to an Fc domain as set forth in SEQ ID NO:163, or a polypeptide comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:163. 
     
     
         86 . A hybrid nuclease molecule comprising a polypeptide, wherein the polypeptide comprises an amino acid sequence of a mature peptide of a human RNase pancreatic precursor operatively linked to an Fc domain via a linker as set forth in SEQ ID NO:161, or a polypeptide comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:161. 
     
     
         87 . A hybrid nuclease molecule comprising a polypeptide, wherein the polypeptide comprises an amino acid sequence of a mature peptide of a human RNase pancreatic precursor operatively linked to the N-terminus of an Fc domain and a human DNase amino acid sequence operatively linked to the C-terminus of the Fc domain as set forth in SEQ ID NO:153, or a polypeptide comprising an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO:153. 
     
     
         88 . The hybrid nuclease molecule of  claim 86 , wherein the Fc domain comprises a modified CH2 domain comprising at least one substitution selected from the group consisting of P238S, P331S, N297S or combination thereof. 
     
     
         89 . A composition comprising the hybrid nuclease molecule of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         90 . A nucleic acid molecule encoding the hybrid nuclease molecule according to  claim 1 . 
     
     
         91 . A recombinant expression vector comprising a nucleic acid molecule according to  claim 90 . 
     
     
         92 . A host cell transformed with the recombinant expression vector according to  claim 91 . 
     
     
         93 . A method of making the hybrid nuclease molecule of  claim 1 , comprising: providing a host cell comprising a nucleic acid sequence that encodes the hybrid nuclease molecule; and maintaining the host cell under conditions in which the hybrid nuclease molecule is expressed. 
     
     
         94 . A method for treating or preventing a condition associated with an abnormal immune response, comprising administering to a subject an effective amount of a hybrid nuclease molecule of  claim 1 . 
     
     
         95 . The method of  claim 94 , wherein the condition is an autoimmune disease. 
     
     
         96 . The method of  claim 95 , wherein the autoimmune disease is selected from the group consisting of insulin-dependent diabetes mellitus, multiple sclerosis, experimental autoimmune encephalomyelitis, rheumatoid arthritis, experimental autoimmune arthritis, myasthenia gravis, thyroiditis, an experimental form of uveoretinitis, Hashimoto's thyroiditis, primary myxoedema, thyrotoxicosis, pernicious anaemia, autoimmune atrophic gastritis, Addison's disease, premature menopause, male infertility, juvenile diabetes, Goodpasture's syndrome, pemphigus vulgaris, pemphigoid, sympathetic ophthalmia, phacogenic uveitis, autoimmune haemolytic anaemia, idiopathic leucopenia, primary biliary cirrhosis, active chronic hepatitis Hbs-ve, cryptogenic cirrhosis, ulcerative colitis, Sjogren's syndrome, scleroderma, Wegener's granulomatosis, polymyositis, dermatomyositis, discoid LE, systemic lupus erythematosus (SLE), and connective tissue disease. 
     
     
         97 . The method of  claim 96 , wherein the autoimmune disease is SLE. 
     
     
         98 . A method of treating SLE comprising administering to a subject a nuclease-containing composition in an amount effective to digest immune complexes containing RNA, DNA or both RNA and DNA, wherein the composition comprises a pharmaceutically acceptable carrier and an isolated polypeptide comprising a first nuclease domain. 
     
     
         99 . The method of  claim 98 , wherein the first nuclease domain comprises an RNase. 
     
     
         100 . The method of  claim 98 , wherein the polypeptide further comprises an Fc domain operatively coupled to the first nuclease domain. 
     
     
         101 . The method of  claim 100 , wherein the Fc domain is modified to decrease binding to Fcγ receptors or complement proteins or both. 
     
     
         102 . The method of  claim 101 , wherein the Fc domain has a reduced effector function selected from the group consisting of opsonization, phagocytosis, complement dependent cytotoxicity, and antibody-dependent cellular cytotoxicity. 
     
     
         103 . The method of  claim 98 , wherein the polypeptide further comprises a second nuclease domain comprising a DNase.