Uses and methods for modulating bile acid homeostasis and treatment of bile acid disorders and diseases

Abstract

The invention relates to variants and fusions of fibroblast growth factor 19 (FGF19), variants and fusions of fibroblast growth factor 21 (FGF21), fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21), and variants or fusions of fibroblast growth factor 19 (FGF19) and/or fibroblast growth factor 21 (FGF21) proteins and peptide sequences (and peptidomimetics), having one or more activities, such as bile acid homeostasis modulating activity, and methods for and uses in treatment of bile acid and other disorders.

1 - 71 . (canceled) 
     
     
         72 . A method of reducing CYP7a1 expression in a subject having nonalcoholic steatohepatitis (NASH), comprising administering to the subject an effective amount of a peptide, wherein the peptide comprises:
 a) an N-terminal region comprising at least seven amino acid residues, the N-terminal region having a first amino acid position and a last amino acid position, wherein the N-terminal region comprises DSSPL (SEQ ID NO:121) or DASPH (SEQ ID NO:122); and   b) a C-terminal region comprising a portion of SEQ ID NO:99 [FGF19], the C-terminal region having a first amino acid position and a last amino acid position, wherein the C-terminal region comprises
 (i) a first C-terminal region sequence comprising WGDPIRLRHLYTSG (amino acids 16 to 29 of SEQ ID NO:99 [FGF19]), wherein the W residue corresponds to the first amino acid position of the C-terminal region; and 
 (ii) a second C-terminal region sequence comprising PHGLSSCFLRIRADGVVDCARGQSAHSLLEIKAVALRTVAIKGVHS VRYLCMGADGKMQGLLQYSEEDCAFEEEIRPD GYNVYRSEKHRL PVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESD MFSSPLETDSMDPFGLVTGLEAVRSPSFEK (amino acid residues 30 to 194 of SEQ ID NO:99 [FGF19]); or a sequence comprising from 1 to 5 amino acid substitutions, deletions or insertions thereof; 
   wherein the peptide   (A) has reduced hepatocellular carcinoma (HCC) formation as compared to FGF19, or as compared to an FGF19 variant sequence having any of GQV, GDI, WGPI (SEQ ID NO:171), WGDPV (SEQ ID NO:172), WGDI (SEQ ID NO:173), GDPI (SEQ ID NO:174), GPI, WGQPI (SEQ ID NO:175), WGAPI (SEQ ID NO:176), AGDPI (SEQ ID NO:177), WADPI (SEQ ID NO:178), WGDAI (SEQ ID NO:179), WGDPA (SEQ ID NO:180), WDPI (SEQ ID NO:181), WGDI (SEQ ID NO:182), WGDP (SEQ ID NO:183) or FGDPI (SEQ ID NO:184), substituted for the WGDPI (SEQ ID NO:170) sequence at amino acids 16-20 of FGF19 (SEQ ID NO:99); and   (B)(i) binds to fibroblast growth factor receptor 4 (FGFR4) with an affinity equal to or greater than FGF19 binding affinity for FGFR4;   (ii) activates FGFR4 to an extent or amount equal to or greater than FGF19 activates FGFR4;   (iii) has at least one of greater glucose lowering activity, less lipid increasing activity, less triglyceride activity, less cholesterol activity, less non-HDL activity or less HDL increasing activity, as compared to FGF19, or as compared to an FGF19 variant sequence having any of GQV, GDI, WGPI (SEQ ID NO:171), WGDPV (SEQ ID NO:172), WGDI (SEQ ID NO:173), GDPI (SEQ ID NO:174), GPI, WGQPI (SEQ ID NO:175), WGAPI (SEQ ID NO:176), AGDPI (SEQ ID NO:177), WADPI (SEQ ID NO:178), WGDAI (SEQ ID NO:179), WGDPA (SEQ ID NO:180), WDPI (SEQ ID NO:181), WGDI (SEQ ID NO:182), WGDP (SEQ ID NO:183) or FGDPI (SEQ ID NO:184), substituted for the WGDPI (SEQ ID NO:170) sequence at amino acids 16-20 of FGF19 (SEQ ID NO:99); and/or   (iv) has less lean mass reducing activity as compared to FGF21;   thereby reducing CYP7a1 expression in said subject without inducing HCC formation.   
     
     
         73 . The method of  claim 72 , wherein the second C-terminal region sequence comprises from 1 to 5 amino acid substitutions, deletions or insertions. 
     
     
         74 . The method of  claim 72 , wherein the peptide is less than about 250 amino acids in length. 
     
     
         75 . The method of  claim 72 , wherein the N-terminal region comprises amino acid residues DASPHVHYG (SEQ ID NO:102), or DSSPLVHYG (SEQ ID NO:103). 
     
     
         76 . The method of  claim 75 , wherein the G corresponds to the last position of the N-terminal region. 
     
     
         77 . The method of  claim 76 , wherein the N-terminal region further comprises:
 RHPIP (SEQ ID NO:106), wherein R is the first amino acid position of the N-terminal region;   HPIP (SEQ ID NO:107), wherein H is the first amino acid position of the N-terminal region;   RPLAF (SEQ ID NO:108), wherein R is the first amino acid position of the N-terminal region;   PLAF (SEQ ID NO:109), wherein P is the first amino acid position of the N-terminal region; or   R, wherein R is the first amino acid position of the N-terminal region.   
     
     
         78 . The method of  claim 72 , wherein the N-terminal region comprises amino acid residues DSSPLLQ (SEQ ID NO:104), and wherein the Q residue is the last amino acid position of the N-terminal region. 
     
     
         79 . The method of  claim 78 , wherein the N-terminal region further comprises:
 RHPIP (SEQ ID NO:106), wherein R is the first amino acid position of the N-terminal region;   HPIP (SEQ ID NO:107), wherein H is the first amino acid position of the N-terminal region;   RPLAF (SEQ ID NO:108), wherein R is the first amino acid position of the N-terminal region;   PLAF (SEQ ID NO:109), wherein P is the first amino acid position of the N-terminal region; or   R, wherein R is the first amino acid position of the N-terminal region.   
     
     
         80 . The method of  claim 72 , wherein the N-terminal region comprises amino acid residues DSSPLLQFGGQV (SEQ ID NO:105), and wherein the V residue corresponds to the last position of the N-terminal region. 
     
     
         81 . The method of  claim 72 , wherein amino acid residues HPIP (SEQ ID NO:107) are the first 4 amino acid residues of the N-terminal region. 
     
     
         82 . The method of  claim 72 , wherein
 the first position of the N-terminal region is a R or M residue;   the first and second positions of the N-terminal region is a MR, RM, RD, DS, MD or MS sequence;   the first through third positions of the N-terminal region is a MDS, RDS, MSD, MSS, or DSS sequence;   the first through fourth positions of the N-terminal region is a RDSS (SEQ ID NO:115) or MDSS (SEQ ID NO:116) sequence;   the first through fifth positions of the N-terminal region is an MRDSS (SEQ ID NO:117) sequence;   the first through sixth positions of the N-terminal region is an MDSSPL (SEQ ID NO:119) sequence; or   the first through seventh positions of the N-terminal region is an MSDSSPL (SEQ ID NO:120) sequence.   
     
     
         83 . The method of  claim 72 , wherein the N-terminal region and the first C-terminal region of the peptide has an amino acid sequence comprising or consisting of any of: 
       
         
           
                 
               
                   RPLAFSDASPHVHYGWGDPIRLRHLYTSG (M1)(amino acids 
                 
                     
                 
                   1-29 of SEQ ID NO: 1); 
                 
                     
                 
                   PLAFSDASPHVHYGWGDPIRLRHLYTSG (M1-R)(amino acids 
                 
                     
                 
                   2-29 of SEQ ID NO: 1); 
                 
                     
                 
                   RPLAFSDSSPLVHYGWGDPIRLRHLYTSG (M2)(amino acids 
                 
                     
                 
                   1-29 of SEQ ID NO: 2); 
                 
                     
                 
                   PLAFSDSSPLVHYGWGDPIRLRHLYTSG (M2-R)(amino acids 
                 
                     
                 
                   2-29 of SEQ ID NO: 2); 
                 
                     
                 
                   RHPIPDSSPLLQWGDPIRLRHLYTSG (M8)(amino acids 1-26 
                 
                     
                 
                   of SEQ ID NO: 8); 
                 
                     
                 
                   RHPIPDSSPLLQFGWGDPIRLRHLYTSG (M9)(amino acids 1-28 
                 
                     
                 
                   of SEQ ID NO: 9); 
                 
                     
                 
                   RPLAFSDSSPLVHWGDPIRLRHLYTSG (M26)(amino acids 1-27 
                 
                     
                 
                   of SEQ ID NO: 26); 
                 
                     
                 
                   PLAFSDSSPLVHWGDPIRLRHLYTSG (M26-R)(amino acids 
                 
                     
                 
                   2-27 of SEQ ID NO: 26); 
                 
                     
                 
                   HPIPDSSPLLQWGDPIRLRHLYTSG (M47)(amino acids 1-25 
                 
                     
                 
                   of SEQ ID NO: 47); 
                 
                     
                 
                   RDSSPLLQWGDPIRLRHLYTSG (M52)(amino acids 1-22 of 
                 
                     
                 
                   SEQ ID NO: 52); 
                 
                     
                 
                   DSSPLLQWGDPIRLRHLYTSG (M52-R)(amino acids 2-22 of 
                 
                     
                 
                   SEQ ID NO: 52); 
                 
                     
                 
                   MDSSPLVHYGWGDPIRLRHLYTSG (M53)(amino acids 1-24 of 
                 
                     
                 
                   SEQ ID NO: 53); 
                 
                     
                 
                   RDSSPLVHYGWGDPIRLRHLYTSG (M69)(amino acids 1-24 of 
                 
                     
                 
                   SEQ ID NO: 69); 
                 
                     
                 
                   DSSPLVHYGWGDPIRLRHLYTSG (M69-R)(amino acids 2-24 
                 
                     
                 
                   of SEQ ID NO: 69); 
                 
                     
                 
                   MRDSSPLVHYGWGDPIRLRHLYTSG (M70)(amino acids 1-25 
                 
                     
                 
                   of SEQ ID NO: 70); 
                 
                     
                 
                   DSSPLVHYGWGDPIRLRHLYTSG (M141)(amino acids 1-23 
                 
                     
                 
                   of SEQ ID NO: 141); 
                 
                   or 
                 
                     
                 
                   HPIPDSSPLLQFGWGDPIRLRHLYTSG (M163)(amino acids 
                 
                     
                 
                   1-27 of SEQ ID NO: 163). 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         84 . The method of  claim 72 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:1 (M1), SEQ ID NO:2 (M2), SEQ ID NO:8 (M8), SEQ ID NO:9 (M9), SEQ ID NO:26 (M26), SEQ ID NO:47 (M47), SEQ ID NO:52 (M52), SEQ ID NO:53, SEQ ID NO:69, (M69), SEQ ID NO:70 (M70), SEQ ID NO:141, SEQ ID NO:163 or SEQ ID NO:192 (M53). 
     
     
         85 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:1. 
     
     
         86 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:2. 
     
     
         87 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:53. 
     
     
         88 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:8. 
     
     
         89 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:9. 
     
     
         90 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:26. 
     
     
         91 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:47. 
     
     
         92 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:141. 
     
     
         93 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:163. 
     
     
         94 . The method of  claim 84 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NO:192. 
     
     
         95 . The method of  claim 72 , wherein the peptide has an amino acid sequence comprising or consisting of SEQ ID NOs:1, 2, 8, 9, 26, 52 or 69, wherein the arginine (R) residue at the first amino acid position of the N-terminal region of the sequence is deleted. 
     
     
         96 . The method of  claim 72 , wherein the N-terminal region first amino acid position is a methionine (M), arginine (R), serine (S), histidine (H), proline (P), leucine (L) or aspartic acid (D) residue. 
     
     
         97 . The method of  claim 72 , wherein the N-terminal region does not have a methionine (M) or arginine (R) residue at the first amino acid position of the N-terminal region. 
     
     
         98 . The method of  claim 72 , wherein the N-terminal region comprises any one of the following amino acid sequences: MDSSPL (SEQ ID NO:119), MSDSSPL (SEQ ID NO:120), or SDSSPL (SEQ ID NO:112). 
     
     
         99 . The method of  claim 72 , wherein the peptide has at least one of greater glucose lowering activity, or less lipid increasing activity as compared to FGF19, or as compared to an FGF19 variant having any of GQV, GDI, WGPI (SEQ ID NO:171), WGDPV (SEQ ID NO:172), WGDI (SEQ ID NO:173), GDPI (SEQ ID NO:174), GPI, WGQPI (SEQ ID NO:175), WGAPI (SEQ ID NO:176), AGDPI (SEQ ID NO:177), WADPI (SEQ ID NO:178), WGDAI (SEQ ID NO:179), WGDPA (SEQ ID NO:180), WDPI (SEQ ID NO:181), WGDI (SEQ ID NO:182), WGDP (SEQ ID NO:183) or FGDPI (SEQ ID NO:184), substituted for the WGDPI (SEQ ID NO:170) sequence at amino acids 16-20 of FGF19 (SEQ ID NO:99). 
     
     
         100 . The method of  claim 72 , wherein the peptide has less lean mass reducing activity as compared to the lean mass reducing activity of FGF21. 
     
     
         101 . The method of  claim 72 , wherein the HCC formation, glucose lowering activity, lipid increasing activity, or lean mass reducing activity is ascertained in a db/db mouse. 
     
     
         102 . The method of  claim 72 , wherein the second C-terminal region sequence comprises a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD sequence (amino acids 74-78 of SEQ ID NO:188). 
     
     
         103 . The method of  claim 72 , wherein the peptide is formulated as a pharmaceutical composition, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. 
     
     
         104 . The method of  claim 72 , wherein the method further comprises administration of a supplemental therapy. 
     
     
         105 . The method of  claim 72 , wherein the peptide is fused with an immunoglobulin Fc region. 
     
     
         106 . A method of reducing CYP7a1 expression in a subject having NASH, comprising administering to the subject an effective amount of a peptide having an amino acid sequence comprising or consisting of
 MRDSSPLVHYGWGDPIRLRHLYTSGPHGLSSCFLRIRADGVVDCARGQSAHSLL EIKAVALRTVAIKGVHSVRYLCMGADGKMQGLLQYSEEDCAFEEEIRPDGYNVY RSEKHRLPVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESDMFS SPLETDSMDPFGLVTGLEAVRSPSFEK (SEQ ID NO:70), or a sequence comprising a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 99-103 of SEQ ID NO:70) sequence thereof,   thereby reducing CYP7a1 expression in said subject.   
     
     
         107 . The method of  claim 106 , wherein the peptide has an amino acid sequence comprising SEQ ID NO:70. 
     
     
         108 . The method of  claim 106 , wherein the peptide has an amino acid sequence consisting of SEQ ID NO:70. 
     
     
         109 . The method of  claim 106 , wherein the peptide is fused with an immunoglobulin Fc region. 
     
     
         110 . The method of  claim 106 , wherein the amino acid sequence comprises a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 99-103 of SEQ ID NO:70) sequence of SEQ ID NO:70. 
     
     
         111 . The method of  claim 106 , wherein the peptide is formulated as a pharmaceutical composition, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. 
     
     
         112 . The method of  claim 106 , wherein the method further comprises administration of a supplemental therapy. 
     
     
         113 . A method of reducing CYP7a1 expression in a subject having NASH, comprising administering to the subject an effective amount of a peptide having an amino acid sequence comprising or consisting of RDSSPLVHYGWGDPIRLRHLYTSGPHGLSSCFLRIRADGVVDCARGQSAHSLLEI KAVALRTVAIKGVHSVRYLCMGADGKMQGLLQYSEEDCAFEEEIRPDGYNVYR SEKHRLPVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESDMFSS PLETDSMDPFGLVTGLEAVRSPSFEK (SEQ ID NO:69), or a sequence comprising a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 98-102 of SEQ ID NO:69) sequence thereof,
 thereby reducing CYP7a1 expression in said subject.   
     
     
         114 . The method of  claim 113 , wherein the peptide has an amino acid sequence comprising SEQ ID NO:69. 
     
     
         115 . The method of  claim 113 , wherein the peptide has an amino acid sequence consisting of SEQ ID NO:69. 
     
     
         116 . The method of  claim 113 , wherein the peptide is fused with an immunoglobulin Fc region. 
     
     
         117 . The method of  claim 113 , wherein the amino acid sequence comprises a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 98-102 of SEQ ID NO:69) sequence of SEQ ID NO:69. 
     
     
         118 . The method of  claim 113 , wherein the peptide is formulated as a pharmaceutical composition, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. 
     
     
         119 . The method of  claim 113 , wherein the method further comprises administration of a supplemental therapy. 
     
     
         120 . A method of reducing CYP7a1 expression in a subject having NASH, comprising administering to the subject an effective amount of a peptide having an amino acid sequence comprising or consisting of
 RDSSPLLQWGDPIRLRHLYTSGPHGLSSCFLRIRADGVVDCARGQ SAHSLLEIKA VALRTVAIKGVHSVRYLCMGADGKMQGLLQYSEEDCAFEEEIRPDGYNVYRSE KHRLPVSLSSAKQRQLYKNRGFLPLSHFLPMLPMVPEEPEDLRGHLESDMFSSPL ETDSMDPFGLVTGLEAVRSPSFEK (SEQ ID NO:52), or a sequence comprising a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 96-100 of SEQ ID NO:52) sequence thereof,   thereby reducing CYP7a1 expression in said subject.   
     
     
         121 . The method of  claim 120 , wherein the peptide has an amino acid sequence comprising SEQ ID NO:52. 
     
     
         122 . The method of  claim 120 , wherein the peptide has an amino acid sequence consisting of SEQ ID NO:52. 
     
     
         123 . The method of  claim 120 , wherein the peptide is fused with an immunoglobulin Fc region. 
     
     
         124 . The method of  claim 120 , wherein the amino acid sequence comprises a EILPD (amino acids 103-107 of SEQ ID NO:193), EIRED (amino acids 103-107 of SEQ ID NO:194), EILCD (amino acids 103-107 of SEQ ID NO:195), EILED (amino acids 103-107 of SEQ ID NO:196), or LLLED (amino acids 98-102 of SEQ ID NO:100) sequence substituted for the EIRPD (amino acids 96-100 of SEQ ID NO:52) sequence of SEQ ID NO:52. 
     
     
         125 . The method of  claim 120 , wherein the peptide is formulated as a pharmaceutical composition, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. 
     
     
         126 . The method of  claim 120 , wherein the method further comprises administration of a supplemental therapy.