RANKL ANTIBODY-PTH/PTHrP CHIMERIC MOLECULES

Abstract

Chimeric molecules comprising receptor activator of NF-κB ligand (RANKL) antibodies and parathyroid hormone/parathyroid hormone-related protein (PTH/PTHrP) peptides are described. Compositions and methods for the treatment of bone diseases are also described.

1 .- 109 . (canceled) 
     
     
         110 . A polynucleotide comprising a polynucleotide encoding a chimeric molecule comprising a heavy chain of a receptor activator of NF-κB ligand (RANKL) antibody and a parathyroid hormone/parathyroid hormone related protein (PTH/PTHrP), wherein the chimeric molecule and a light chain of the RANKL antibody (i) inhibit binding of RANKL to a receptor activator of NF-κB (RANK) and (ii) bind to a PTH-1 receptor and/or a PTH-2 receptor. 
     
     
         111 . The polynucleotide of  claim 110 , wherein the heavy chain comprises an amino acid sequence that has at least 92% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         112 . The polynucleotide of  claim 110 , wherein the light chain comprises an amino acid sequence that has at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         113 . The polynucleotide of  claim 110 , wherein the heavy chain comprises an amino acid sequence that has at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         114 . The polynucleotide of  claim 110 , wherein the light chain comprises an amino acid sequence that has at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         115 . The polynucleotide of  claim 110 , the heavy chain comprises an amino acid sequence that has at least 99% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         116 . The polynucleotide of  claim 110 , wherein the light chain comprises an amino acid sequence that has at least 99% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         117 . The polynucleotide of  claim 110 , wherein the heavy chain comprises an amino acid sequence as set forth in SEQ ID NO: 11. 
     
     
         118 . The polynucleotide of  claim 110 , wherein the light chain comprises an amino acid sequence as set forth in SEQ ID NO: 12. 
     
     
         119 . The polynucleotide of  claim 110  the heavy chain comprises an amino acid sequence as set forth in SEQ ID NO: 2 from residue 20 to residue 467, and the light chain comprises an amino acid sequence as set forth in SEQ ID NO: 4 from residue 21 to residue 235. 
     
     
         120 . The polynucleotide of  claim 110 , wherein the PTH/PTHrP peptide comprises a PTH/PTHrP modulating domain selected from the polypeptides of the following formula:
   X N X 8 HX 10 X 11 X 12 KX 14 X 15 X 16 X 17 X 18 X 19 R 21 X 22 X 23 X 24 X 25 X 26 X 27 X 28 X C      wherein:
 X N X 1 X 2 X 3 X 4 X 5 X 6 X 7 ; 
 X 1  is A, S or Y; 
 X 2  is V; 
 X 3  is S; 
 X 4  is E; 
 X 5  is H or I; 
 X 6  is Q or E; 
 X 7  is L or F; 
 X 8  is M; 
 X 10  is N or D; 
 X 11  is L, R, or K; 
 X 12  is G, F, or W; 
 X 14  is H or S; 
 X 15  is L or I; 
 X 16  is Q, N, S, or A; 
 X 17  is S, D, or L; 
 X 18  is M, L, or V; 
 X 19  is E or R; 
 X 21  is V, M, or R; 
 X 22  is E or F; 
 X 23  is W or F; 
 X 23  is W or F; 
 X 24  is L; 
 X 25  is R or H; 
 X 26  is K or H; 
 X 27  is K or L; 
 X 28  is L or I; 
 X C  is X 29 X 30 X 31 X 32 X 33 X 34 ; 
 X 29  is Q or A; 
 X 30  is D or E; 
 X 31  is V or I; 
 X 33  is N or T; and 
 X 34  is A, F or Y. 
   
     
     
         121 . The polynucleotide of  claim 110 , wherein the PTH/PTHrP peptide comprises a PTH/PTHrP modulating domain, wherein the PTH/PTHrP modulating domain comprises:
 a) a sequence selected from SEQ ID NOs: 16 to 89; or   b) a sequence selected from SEQ ID NOs: 90 to 107 except that one or more residues at position 14 through the C-terminus of the PTH/PTHrP modulating domain is substituted with a cysteine residue.   
     
     
         122 . The polynucleotide of  claim 110 , wherein the PTH/PTHrP peptide comprises a prepro domain and a PTH/PTHrP modulating domain, wherein the prepro domain comprises an amino acid sequence selected from SEQ ID NOs: 188 to 207. 
     
     
         123 . The polynucleotide of  claim 122 , wherein the prepro domain comprises the amino acid sequence of SEQ ID NO: 188 and the PTH/PTHrP modulating domain comprises the amino acid sequence of SEQ ID NO: 22. 
     
     
         124 . The polynucleotide of  claim 110 , wherein the PTH/PTHrP peptide comprises an amino acid sequence of SEQ ID NO: 6 from residue 32 to residue 70. 
     
     
         125 . The polynucleotide of  claim 110 , wherein the chimeric molecule comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 10 from residue 32 to residue 518. 
     
     
         126 . A polynucleotide comprising a polynucleotide encoding a chimeric molecule comprising a light chain of a receptor activator of NF-κB ligand (RANKL) antibody and a parathyroid hormone/parathyroid hormone related protein (PTH/PTHrP), wherein the chimeric molecule and a heavy chain of the RANKL antibody (i) inhibit binding of RANKL to a receptor activator of NF-κB (RANK) and (ii) bind to a PTH-1 receptor and/or a PTH-2 receptor. 
     
     
         127 . The polynucleotide of  claim 126 , wherein the heavy chain comprises an amino acid sequence that has at least 92% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         128 . The polynucleotide of  claim 126 , wherein the light chain comprises an amino acid sequence that has at least 90% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         129 . The polynucleotide of  claim 126 , wherein the heavy chain comprises an amino acid sequence that has at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         130 . The polynucleotide of  claim 126 , wherein the light chain comprises an amino acid sequence that has at least 95% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         131 . The polynucleotide of  claim 126 , the heavy chain comprises an amino acid sequence that has at least 99% identity to the amino acid sequence set forth in SEQ ID NO: 11. 
     
     
         132 . The polynucleotide of  claim 126 , wherein the light chain comprises an amino acid sequence that has at least 99% identity to the amino acid sequence set forth in SEQ ID NO: 12. 
     
     
         133 . The polynucleotide of  claim 126 , wherein the heavy chain comprises an amino acid sequence as set forth in SEQ ID NO: 11. 
     
     
         134 . The polynucleotide of  claim 126 , wherein the light chain comprises an amino acid sequence as set forth in SEQ ID NO: 12. 
     
     
         135 . The polynucleotide of  claim 126  the heavy chain comprises an amino acid sequence as set forth in SEQ ID NO: 2 from residue 20 to residue 467, and the light chain comprises an amino acid sequence as set forth in SEQ ID NO: 4 from residue 21 to residue 235. 
     
     
         136 . The polynucleotide of  claim 126 , wherein the PTH/PTHrP peptide comprises a PTH/PTHrP modulating domain selected from the polypeptides of the following formula:
   X N X 8 HX 10 X 11 X 12 KX 14 X 15 X 16 X 17 X 18 X 19 RX 21 X 22 X 23 X 24 X 25 X 26 X 27 X 28 X C      wherein:
 X N X 1 X 2 X 3 X 4 X 5 X 6 X 7 ; 
 X 1  is A, S or Y; 
 X 2  is V; 
 X 3  is S; 
 X 4  is E; 
 X 5  is H or I; 
 X 6  is Q or E; 
 X 7  is L or F; 
 X 8  is M; 
 X 10  is N or D; 
 X 11  is L, R, or K; 
 X 12  is G, F, or W; 
 X 14  is H or S; 
 X 15  is L or I; 
 X 16  is Q, N, S, or A; 
 X 17  is S, D, or L; 
 X 18  is M, L, or V; 
 X 19  is E or R; 
 X 21  is V, M, or R; 
 X 22  is E or F; 
 X 23  is W or F; 
 X 23  is W or F; 
 X 24  is L; 
 X 25  is R or H; 
 X 26  is K or H; 
 X 27  is K or L; 
 X 28  is L or I; 
 X C  is X 29 X 30 X 31 X 32 X 33 X 34 ; 
 X 29  is Q or A; 
 X 30  is D or E; 
 X 31  is V or I; 
 X 33  is N or T; and 
 X 34  is A, F or Y. 
   
     
     
         137 . The polynucleotide of  claim 126 , wherein the PTH/PTHrP peptide comprises a PTH/PTHrP modulating domain, wherein the PTH/PTHrP modulating domain comprises:
 a) a sequence selected from SEQ ID NOs: 16 to 89; or   b) a sequence selected from SEQ ID NOs: 90 to 107 except that one or more residues at position 14 through the C-terminus of the PTH/PTHrP modulating domain is substituted with a cysteine residue.   
     
     
         138 . The polynucleotide of  claim 126 , wherein the PTH/PTHrP peptide comprises a prepro domain and a PTH/PTHrP modulating domain, wherein the prepro domain comprises an amino acid sequence selected from SEQ ID NOs: 188 to 207. 
     
     
         139 . The polynucleotide of  claim 138 , wherein the prepro domain comprises the amino acid sequence of SEQ ID NO: 188 and the PTH/PTHrP modulating domain comprises the amino acid sequence of SEQ ID NO: 22. 
     
     
         140 . The polynucleotide of  claim 126 , wherein the PTH/PTHrP peptide comprises an amino acid sequence of SEQ ID NO: 6 from residue 32 to residue 70. 
     
     
         141 . The polynucleotide of  claim 126 , wherein the chimeric molecule comprises a polypeptide comprising an amino acid sequence of SEQ ID NO: 8 from residue 32 to residue 285. 
     
     
         142 . A vector comprising a polynucleotide encoding a chimeric molecule of  claim 110  or  claim 126 . 
     
     
         143 . A host cell comprising the vector of  claim 142 . 
     
     
         144 . A method of producing a chimeric molecule comprising culturing the host cell of  claim 143 .