Fgf21 variants

Patent metadata

Field	Value
Publication number	US-2025171512-A1
Application number	US-202418932818-A
Country	US
Kind code	A1
Filing date	Oct 31, 2024
Priority date	Dec 2, 2015
Publication date	May 29, 2025
Grant date	—

Abstract

The present invention relates to polypeptide variants of human fibroblast growth factor 21 (FGF21) and fusion molecules thereof, as well as to nucleic acid molecules encoding the same. It further relates to their use as medicaments, in particular for the treatment of obesity, overweight, metabolic syndrome, diabetes mellitus, hyperglycemia, dyslipidemia, non-alcoholic steatohepatitis (NASH) and/or atherosclerosis.

First claim

1 . A variant of human fibroblast growth factor 21 (FGF21) comprising the amino acid sequence of SEQ ID NO: 171,
wherein
Xaa55 is Q or C;
Xaa147 is P or C;
Xaa148 is G or C;
Xaa149 is N or C;
Xaa150 is K;
Xaa151 is S;
Xaa152 is P;
Xaa153 is H;
Xaa154 is R;
Xaa155 is D or C;
Xaa156 is P;
Xaa157 is A;
Xaa158 is P;
Xaa159 is R;
Xaa160 is G;
Xaa161 is P or C;
Xaa162 is A or C;
Xaa163 is R;
Xaa184 is Q;
Xaa185 is P;
Xaa186 is P;
Xaa187 is D;
Xaa188 is V;
Xaa189 is G;
Xaa190 is S;
Xaa191 is S;
Xaa192 is D;
Xaa193 is P;
Xaa194 is L;
Xaa195 is S or C;
Xaa196 is M or another amino acid, such as P, V or C, or deleted;
Xaa197 is V or another amino acid, such as E, D, G or M, or deleted;
Xaa198 is G or another amino acid, such as E, D, R, K, Y, P or V, or deleted;
Xaa199 is P or another amino acid, such as S, Q, R, T, G, F, L, D or M, or deleted;
Xaa200 is S or another amino acid, such as Q, M, C, P, N or H, or deleted;
Xaa201 is Q or another amino acid, such as P or S, or deleted;
Xaa202 is G or another amino acid, such as T, or deleted;
Xaa203 is R or another amino acid, such as E, H or C, or deleted;
Xaa204 is S;
Xaa205 is P;
Xaa206 is S; and
Xaa207 is Y;
wherein, optionally, SEQ ID NO: 171 comprises a substitution of at least one of the following amino acids with C: R47, L49, T51, A54, Q56, A59, H60, E62, 163, G67, V69, G71, A72, A73, S76, P77, E78, S79, L80, L81, Q82, L83, 191, L94, G95, V96, K97, T98, R100, L102, Q104, D107, G108, L110, G112, L114, A120, R124, D130, Y132, Q136, S137, A139, H140, L142, P143, H145, L146, L165, L167, L170, P174; with the proviso that SEQ ID NO: 171 is not mature human wild-type FGF21 (SEQ ID NO: 2) and comprises 0, 2, 4, 6 or 8 additional cysteines as compared to mature human wild-type FGF21 (SEQ ID NO: 2), wherein, optionally, SEQ ID NO: 171 further comprises the mutation G141S and/or the mutation P174L.

2 . The variant according to claim 1 comprising the amino acid sequence of SEQ ID NO: 175.

3 . The variant according to claim 1 , wherein
Xaa197 to Xaa203 are deleted and replaced by a protease resistant peptide linker, such as selected from SEQ ID NO: 166 and SEQ ID NO: 167; a protease resistant peptide linker, such as SEQ ID NO: 168, is inserted between Xaa198 and Xaa199; and/or the amino acid sequence of SEQ ID NO: 169 or SEQ ID NO: 170 is added after S209 of SEQ ID NO: 171.

4 . The variant according to claim 1 , wherein SEQ ID NO: 171 has at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% sequence identity with mature human wild-type FGF21 (SEQ ID NO: 2).

5 . The variant according to claim 1 , wherein
Xaa55 is Q or C; Xaa147 is P or C; Xaa148 is G; Xaa149 is N or C; Xaa155 is D or C; Xaa161 is P; Xaa162 is A; Xaa195 is S; Xaa196 is M; Xaa197 is V; Xaa198 is G or another amino acid, such as E, D, R, K, Y, P or V, or deleted; Xaa199 is P or another amino acid, such as S, Q, R, T, G, F, L, D or M, or deleted; Xaa200 is S; Xaa201 is Q; Xaa202 is G; and Xaa203 is R;
and wherein, optionally, SEQ ID NO: 171 comprises a substitution of at least one of the following amino acids with C: R47, A59, H60, G71, S76, S79, D107, G108, L142, P174.

6 . The variant according to claim 5 comprising the amino acid sequence of SEQ ID NO: 176.

7 . The variant according to claim 1 , wherein SEQ ID NO: 171 comprises a substitution of any of the following amino acids with C:
Xaa55 and Xaa149; R47 and P174; Xaa55 and Xaa147; A59 and G71; H60 and S79; S76 and S79; D107 and Xaa155; and/or G108 and L142.

8 . The variant according to claim 1 , wherein
Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted; Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P; Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted; Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is Y, and Xaa199 is P; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is R, and Xaa199 is P; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is K, and Xaa199 is P; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P; Xaa55 is C, Xaa147 is C, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is P; Xaa55 is C, Xaa147 is P, Xaa149 is C, Xaa155 is D, Xaa198 is G, and Xaa199 is P; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171 comprises the substitution of A59 and G71 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171 comprises the substitution of A59 and G71 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171 comprises the substitution of S76 and S79 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171 comprises the substitution of S76 and S79 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171 comprises the substitution of G108 and L142 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is D, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171 comprises the substitution of G108 and L142 with C; Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is C, Xaa198 is G, and Xaa199 is deleted, wherein SEQ ID NO: 171 comprises the substitution of D107 with C; or Xaa55 is Q, Xaa147 is P, Xaa149 is N, Xaa155 is C, Xaa198 is Y, and Xaa199 is P, wherein SEQ ID NO: 171 comprises the substitution of D107 with C.

9 . The variant according to claim 1 , further comprising at least one additional amino acid at its N-terminus, optionally wherein the at least one additional amino acid is selected from the group consisting of G, A, N and C.

10 . (canceled)

11 . A variant of human FGF21 comprising or consisting of an amino acid sequence according to one of SEQ ID NOs: 269, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 61, 62, 63, 64, 69, 70, 71, 72, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 153, 154, 155, 156, 157, 158, 159 160, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 230, 231, 232, 233, 238, 239, 240, 241, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 268, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 322, 323, 324, 325, 326, 327 and 328, wherein, optionally, the variant further comprises the mutation G141S and/or the mutation P174L, wherein the numbering of the amino acids is in accordance with SEQ ID NO: 1.

12 . The variant according to claim 1 , wherein the variant:
has an increased proteolytic stability in human and/or murine blood plasma as compared to mature human wild-type FGF21 (SEQ ID NO: 2); has an increased thermal stability as compared to mature human wild-type FGF21 (SEQ ID NO: 2); induces phosphorylation of the mitogen-activated protein kinase (MAPK) ERK1/2; or induces phosphory

Assignees

• Sanofi Sa

Inventors

• Sommerfeld Mark
• Langer Thomas
• Boscheinen Oliver
• Dreyer Matthias
• Dittrich Werner
• Habermann Paul

Classifications

• C07K2319/31

  fusions, other than Fc, for prolonged plasma life, e.g. albumin · CPC title

• C07K2319/30

  Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto · CPC title

• C07K2319/21

  containing a His-tag · CPC title

• C07K2319/20

  containing a tag with affinity for a non-protein ligand · CPC title

• C07K14/78

  Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG] · CPC title