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, a second nuclease domain, and an Fc domain, wherein the Fc domain is operatively coupled to either the first or second nuclease domain. 
     
     
         2 . The hybrid nuclease molecule of  claim 1 , wherein the first and second nuclease domains are distinct nuclease domains. 
     
     
         3 . The hybrid nuclease molecule of  claim 1 , wherein the first and second nuclease domains are the same nuclease domains. 
     
     
         4 . The hybrid nuclease molecule of  claim 1 , wherein the first nuclease domain is operatively coupled to the Fc domain, and wherein the Fc domain is operatively coupled to the second nuclease domain. 
     
     
         5 . The hybrid nuclease molecule of  claim 4 , wherein the first nuclease domain is linked to the N-terminus of the Fc domain and the second nuclease domain is linked to the C-terminus of the Fc domain. 
     
     
         6 . The hybrid nuclease molecule of  claim 4 , wherein the first nuclease domain is linked to the C-terminus of the Fc domain and the second nuclease domain is linked to the N-terminus of the Fc domain. 
     
     
         7 . The hybrid nuclease molecule of  claim 1 , wherein the second nuclease domain is linked to the C-terminus of the first nuclease domain and the Fc domain is linked to the N-terminus of the first nuclease domain. 
     
     
         8 . The hybrid nuclease molecule of  claim 1 , wherein the second nuclease domain is linked to the N-terminus of the first nuclease domain and the Fc domain is linked to the C-terminus of the first nuclease domain. 
     
     
         9 . The hybrid nuclease molecule of  claim 1 , wherein the Fc domain is a mutant Fc domain. 
     
     
         10 . The hybrid nuclease of  claim 9 , wherein the Fc domain comprises a human immunoglobulin Fc domain. 
     
     
         11 . The hybrid nuclease molecule of  claim 10 , wherein the human immunoglobulin Fc domain is a human IgG1 Fc domain. 
     
     
         12 . The hybrid nuclease molecule of  claim 11 , wherein the Fc domain comprises a hinge domain, a CH2 domain and a CH3 domain. 
     
     
         13 . The hybrid nuclease molecule of  claim 12 , wherein the Fc domain comprises a modified hinge domain comprising at least one substitution. 
     
     
         14 . The hybrid nuclease molecule of  claim 13 , wherein the modified hinge domain comprises at least one cysteine substitution. 
     
     
         15 . The hybrid nuclease molecule of  claim 14 , wherein the cysteine is substituted with serine. 
     
     
         16 . The hybrid nuclease molecule of  claim 12 , wherein the Fc domain comprises a modified CH2 domain comprising at least one substitution. 
     
     
         17 . The hybrid nuclease molecule of  claim 16 , wherein the modified CH2 domain comprises at least one substitution of an amino acid associated with Fcγ receptor binding, complement protein binding, or both Fcγ receptor binding and complement protein binding. 
     
     
         18 . The hybrid nuclease molecule of  claim 17 , wherein the substitution is selected from the group consisting of P238S, P331S, N297S, and combinations thereof. 
     
     
         19 . The hybrid nuclease molecule of  claim 9 , wherein the mutant Fc domain has decreased binding to Fcγ receptors, complement proteins, or both Fcγ receptors and complement proteins. 
     
     
         20 . The hybrid nuclease molecule of  claim 9 , wherein the hybrid nuclease molecule has a reduced effector function selected from the group consisting of opsonization, phagocytosis, complement dependent cytotoxicity, and antibody-dependent cellular cytotoxicity. 
     
     
         21 . The hybrid nuclease molecule of  claim 1 , wherein the first nuclease domain comprises a DNase. 
     
     
         22 . The hybrid nuclease molecule of  claim 21 , wherein the DNase is selected from the group consisting of a Type I human DNase, a human DNase 1L3, and human TREX1. 
     
     
         23 . The hybrid nuclease molecule of  claim 22 , wherein the DNase is human DNase1. 
     
     
         24 . The hybrid nuclease molecule of  claim 1 , wherein the second nuclease domain comprises an RNase. 
     
     
         25 . The hybrid nuclease molecule of  claim 24 , wherein the RNase is a human RNase. 
     
     
         26 . The hybrid nuclease molecule of  claim 25 , wherein the human RNase is human pancreatic RNase A. 
     
     
         27 . The hybrid nuclease molecule of  claim 1 , wherein the hybrid nuclease molecule further comprises a first linker domain, a second linker domain, or both, wherein when the molecule comprises a first linker domain, the first nuclease domain is operatively coupled to the Fc domain by the first linker domain, and when the molecule comprises a second linker domain, the second nuclease domain is operatively coupled to the Fc domain by the second linker domain. 
     
     
         28 . The hybrid nuclease molecule of  claim 27 , wherein the hybrid nuclease molecule further comprises a first linker domain and a second linker domain, wherein the amino acid sequence of the first nuclease domain comprises a human DNase1 amino acid sequence, wherein the amino acid sequence of the Fc domain comprises a mutant human Fc domain amino acid sequence, and wherein the amino acid sequence of the second nuclease domain comprises a human RNase1 amino acid sequence. 
     
     
         29 . The hybrid nuclease molecule of  claim 28 , wherein the first linker domain is a gly/ser peptide, wherein the gly/ser peptide is of the formula (Gly4Ser)n, wherein n is a positive integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, and the second linker domain is an NLG peptide between 5 and 32 amino acids in length, wherein the first linker domain is coupled to the C-terminus of the first nuclease domain and the N-terminus of the mutant human Fc domain, and wherein the second linker domain is coupled to the C-terminus of the mutant human Fc domain and the N-terminus of the second nuclease domain. 
     
     
         30 . The hybrid nuclease molecule of  claim 1 , wherein the hybrid nuclease molecule further comprises a first linker domain and a second linker domain, wherein the amino acid sequence of the first nuclease domain comprises a human RNase1 amino acid sequence, wherein the amino acid sequence of the Fc domain comprises a mutant human Fc domain amino acid sequence, and wherein the amino acid sequence of the second nuclease domain comprises a human DNase1 amino acid sequence. 
     
     
         31 . The hybrid nuclease molecule of  claim 30 , wherein the first linker domain is a gly/ser peptide, wherein the gly/ser peptide is of the formula (Gly4Ser)n, wherein n is a positive integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10, and the second linker domain is an NLG peptide between 5 and 32 amino acids in length, wherein the first linker domain is coupled to the C-terminus of the first nuclease domain and the N-terminus of the mutant human Fc domain, and wherein the second linker domain is coupled to the C-terminus of the mutant human Fc domain and the N-terminus of the second nuclease domain. 
     
     
         32 . The hybrid nuclease molecule of  claim 1 , 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:149. 
     
     
         33 . 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:145. 
     
     
         34 . The hybrid nuclease molecule of  claim 33 , wherein the Fc domain further comprises one or more mutations selected from the group consisting of P238S, P331S, K322S, N297S, SCC, SSS, and combinations thereof. 
     
     
         35 . The hybrid nuclease molecule of  claim 34 , wherein the mutation is P238S. 
     
     
         36 . The hybrid nuclease molecule of  claim 1 , 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:143. 
     
     
         37 . The hybrid nuclease molecule of  claim 1 , wherein the second nuclease domain comprises a human DNase comprising an amino acid sequence set forth in SEQ ID NO:142, or a DNase comprising an amino acid sequence at least 90% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:142. 
     
     
         38 . The hybrid nuclease molecule of  claim 1 , 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:149, 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:145, and 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% or at least 95% identical to the amino acid sequence set forth in SEQ ID NO:143. 
     
     
         39 . The hybrid nuclease molecule of  claim 38 , wherein the Fc domain further comprises one or more mutations selected from the group consisting of P238S, P331S, K322S, N297S, SCC, SSS, and combinations thereof. 
     
     
         40 . The hybrid nuclease molecule of  claim 39 , wherein the mutation is P238S. 
     
     
         41 . A hybrid nuclease molecule comprising 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. 
     
     
         42 . The hybrid nuclease molecule of  claim 41 , wherein the Fc domain further comprises one or more mutations selected from the group consisting of P238S, P331S, K322S, N297S, SCC, SSS, and combinations thereof. 
     
     
         43 . The hybrid nuclease molecule of  claim 42 , wherein the mutation is P238S. 
     
     
         44 . A composition comprising the hybrid nuclease molecule of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         45 . A method for treating or preventing a condition associated with an abnormal immune response, comprising administering to a subject an effective amount of a the composition of  claim 47 . 
     
     
         46 . The method of  claim 45 , wherein the condition is an autoimmune disease. 
     
     
         47 . The method of  claim 46 , 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. 
     
     
         48 . The method of  claim 47 , wherein the autoimmune disease is SLE.