Engineered Polypeptides Having Enhanced Duration of Action and Reduced Immunogenicity

Abstract

Compounds are provided having inter alia good duration of action, high potency and/or convenient dosing regimens including once weekly administration. The compounds are engineered polypeptides which incorporate an albumin binding domain in combination with one or more biologically active polypeptides. Also provided are pharmaceutical compositions and methods of treatment for diseases and disorders including lipodystrophy, dyslipidemia, hyperlipidemia, overweight, obesity, hypothalamic amenorrhea, Alzheimer's disease, leptin deficiency, fatty liver disease or diabetes (including type I and type II). Additional diseases and disorders which can be treated by the compounds and methods described herein include nonalcoholic steatohepatitis (NASH) and nonalcoholic fatty liver disease (NAFLD), metabolic syndrome X and Huntington's Disease.

1 . An engineered polypeptide comprising: an albumin binding domain polypeptide (ABD) and a first peptide hormone domain (HD1) selected from a leptin, a leptin analog or an active fragment thereof,
 wherein said ABD comprises an amino acid sequence selected from the amino acid sequence comprising:   (i) LA X3 AK X6 X7 AN X10 ELD X14 YGVSDF YKRL1 X26 KAKT VEGVEALK X39 X40 IL X43 X44 LP (SEQ ID NO: 300)   
       wherein independently of each other 
       X3 is selected from E, S, Q and C; 
       X6 is selected from E, S and C; 
       X7 is selected from A and S; 
       X10 is selected from A, S and R; 
       X14 is selected from A, S, C and K; 
       X26 is selected from D and E; 
       X39 is selected from D and E; 
       X40 is selected from A and E; 
       X43 is selected from A and K; 
       X44 is selected from A, S and E; 
       the leucine at position 45 is present or absent; and 
       the proline at position 46 is present or absent; and
 (ii) an amino acid sequence which has at least 95% identity to the sequence defined in (i); 
 with the proviso that X7 is not L, E or D; 
 or alternatively, 
 with the proviso that the amino acid sequence is not defined by the following sequence, as defined in PCT Published Application No. WO 2009/016043: LAEAK X a  X b  A X c  X d  EL X e  KY GVSD X 5  YK X 8  X 9  I X 11  X 12  A X 14  TVEGV X 20  AL X 23  X 24  X 25  ILAALP (SEQ ID NO: 679) 
 
       wherein
 independently of each other, 
 X a  is selected from V and E; 
 X b  is selected from L, E and D; 
 X c  is selected from N, L and I; 
 X d  is selected from R and K; 
 X e  is selected from D and K; and 
 X 5  is selected from Y and F; 
 X 8  is selected from N, R and S; 
 X 9  is selected from V, I, L, M, F and Y; 
 X 11  is selected from N, S, E and D; 
 X 12  is selected from R, K and N; 
 X 14  is selected from K and R; 
 X 20  is selected from D, N, Q, E, H, S, R and K; 
 X 23  is selected from K, I and T; 
 X 24  is selected from A, S, T, G, H, L and D; and 
 X 25  is selected from H, E and D. 
 
     
     
         2 . The engineered polypeptide according to  claim 1 , further comprising a first linker (L1) covalently linked to said HD1. 
     
     
         3 . The engineered polypeptide according to  claim 1 , wherein said engineered polypeptide comprises said ABD as an N-terminal moiety and said HD1 as a C-terminal moiety. 
     
     
         4 . The engineered polypeptide according to  claim 1 , wherein said engineered polypeptide comprises said ABD as a C-terminal moiety and said HD1 as an N-terminal moiety. 
     
     
         5 - 8 . (canceled) 
     
     
         9 . The engineered polypeptide according to  claim 1 , wherein said HD1 is a leptin, a leptin analog, a leptin active fragment, or a leptin derivative. 
     
     
         10 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 50% identity with an amino acid sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO: 143, SEQ TD NO: 144, SEQ TD NO: 145, SEQ TD NO: 146, SEQ TD NO:664, SEQ TD 20 NO:665, SEQ ID NO:666, SEQ ID NO:667, SEQ ID NO:668, SEQ ID NO:669, SEQ ID NO:670, SEQ ID NO:671, SEQ ID NO:672, SEQ ID NO:673, SEQ ID NO:674, SEQ ID NO:675, SEQ ID NO:676, SEQ ID NO:677, and SEQ ID NO:680. 
     
     
         11 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 90% identity with an amino acid sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO: 143, SEQ TD NO: 144, SEQ TD NO: 145, SEQ TD NO: 146, SEQ TD NO:664, SEQ TD 20 NO:665, SEQ ID NO:666, SEQ ID NO:667, SEQ ID NO:668, SEQ ID NO:669, SEQ ID NO:670, SEQ ID NO:671, SEQ ID 6 of 12 NO:672, SEQ ID NO:673, SEQ ID NO:674, SEQ ID NO:675, SEQ ID NO:676, SEQ ID NO:677, and SEQ ID NO:680. 
     
     
         12 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 50% identity with a human leptin. 
     
     
         13 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 90% identity with a human leptin. 
     
     
         14 - 15 . (canceled) 
     
     
         16 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 50% identity with a platypus leptin. 
     
     
         17 . The engineered polypeptide according to  claim 1 , wherein said HD1 has at least 50% identity with a seal leptin. 
     
     
         18 . The engineered polypeptide according to  claim 1 , wherein said HD1 has from 1 to 5 amino acid modifications selected independently from any one or combination of an insertion, deletion, addition and substitution. 
     
     
         19 - 74 . (canceled) 
     
     
         75 . The engineered polypeptide according to  claim 1 , wherein the ABD is selected from any one of SEQ ID NO:301-452, 455-463, and 500-593. 
     
     
         76 . The engineered polypeptide according to  claim 1 , wherein the ABD is selected from an amino acid sequence selected from any one of SEQ ID NO:313, SEQ ID NO:448, SEQ ID NO:463, SEQ ID NO:500, SEQ ID NO:501, and SEQ ID NO:502. 
     
     
         77 - 85 . (canceled) 
     
     
         86 . The engineered polypeptide according  claim 1 , wherein the HD1 is conjugated to the ABD via a thiol group of a cysteine residue at position X 14  of the polypeptide. 
     
     
         87 . The engineered polypeptide according to  claim 2 , wherein said linker L1 is a peptide of from 1 to 30 amino acids or less than 30 amino acids. 
     
     
         88 - 92 . (canceled) 
     
     
         93 . The engineered polypeptide according to  claim 2 , wherein said linker L1 comprises polyglycine, polyalanines, poly(Gly-Ala), or poly(Gly-Ser). 
     
     
         94 - 190 . (canceled) 
     
     
         191 . A method for treating a disease or disorder in a subject, comprising administering a engineered polypeptide according to  claim 1  to a subject in need thereof in an amount effective to treat said disease or disorder. 
     
     
         192 . The method according to  claim 191 , wherein said disease or disorder is disease or disorder can be lipodystrophy, dyslipidemia, hyperlipidemia, overweight, obesity, hypothalamic amenorrhea, Alzheimer's disease, leptin deficiency, fatty liver disease, diabetes (including type I and type II), nonalcoholic steatohepatitis (NASH), nonalcoholic fatty liver disease (NAFLD), metabolic syndrome X and Huntington's Disease. 
     
     
         193 - 196 . (canceled) 
     
     
         197 . A pharmaceutical composition comprising an engineered polypeptide according to  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         198 - 283 . (canceled)