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BR-122026002676-A2 - Antibody and its uses, isolated polynucleotide, vector, pharmaceutical composition, method of producing the antibody or a fragment thereof.

BR122026002676A2BR 122026002676 A2BR122026002676 A2BR 122026002676A2BR-122026002676-A2

Abstract

The present disclosure provides binding proteins, such as antibodies, that bind to a GDNF family receptor-type alpha protein (GFRAL), including human GFRAL protein, and methods for their use.

Inventors

  • Wenyan Shen
  • Jie Tang
  • Yan Wang
  • Hugo Matern

Assignees

  • NGM Biopharmaceuticals, Inc

Dates

Publication Date
20260317
Application Date
20170303
Priority Date
20160331

Claims (11)

  1. 1. An antibody or fragment thereof that binds to a GDNF family receptor-type alpha protein (GFRAL), characterized in that the antibody or fragment thereof comprises a variable heavy chain (VH) region and a variable light chain (VL) region wherein: i. the VH region comprises a complementary determinant region (CDR)1, a CDR2 and a CDR3 comprising the amino acid sequences set forth in SEQ ID NOs: 46, 137 and 225, respectively; and ii. the VL region comprises a CDR1, a CDR2 and a CDR3 comprising the amino acid sequences set forth in SEQ ID NOs: 301, 376 and 426, respectively.
  2. 2. An antibody or fragment thereof that binds to a GDNF family receptor-type alpha protein (GFRAL), characterized in that the antibody or fragment thereof comprises a variable heavy chain (VH) region comprising the amino acid sequence set forth in SEQ ID NO: 1982 and a variable light chain (VL) region comprising the amino acid sequence set forth in SEQ ID NO: 1997.
  3. 3. An antibody or fragment thereof characterized in that it competes for binding to human GFRAL with an antibody or fragment thereof, as defined in claim 1.
  4. 4. An antibody or fragment thereof that binds to a human GDNF family receptor-type alpha protein (GFRAL), characterized in that it comprises: a. a variable heavy chain (VH) region comprising a variable heavy chain (VH) region complementary to the determining region (CDR)1, a CDR2 of VH and a CDR3 of VH comprising the amino acid sequences set forth in SEQ ID NO: 56, SEQ ID NO: 147 and SEQ ID NO: 233, respectively; and a variable light chain (VL) region comprising a CDR1 of VL, a CDR2 of VL and a CDR3 of VL comprising the amino acid sequences set forth in SEQ ID NO: 309, SEQ ID NO: 382 and SEQ ID NO: 432, respectively; b. a. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 86, SEQ ID NO: 177, and SEQ ID NO: 261, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 337, SEQ ID NO: 401, and SEQ ID NO: 453, respectively; c. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 91, SEQ ID NO: 182, and SEQ ID NO: 265, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 341, SEQ ID NO: 385, and SEQ ID NO: 456, respectively; d. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 95, SEQ ID NO: 187, and SEQ ID NO: 269, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 345, SEQ ID NO: 404, and SEQ ID NO: 459, respectively; e. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 124, SEQ ID NO: 210, and SEQ ID NO: 289, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 365, SEQ ID NO: 418, and SEQ ID NO: 474, respectively; f. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 127, SEQ ID NO: 215, and SEQ ID NO: 293, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 369, SEQ ID NO: 421, and SEQ ID NO: 477, respectively; g. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 41, SEQ ID NO: 132, and SEQ ID NO: 221, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 297, SEQ ID NO: 373, and SEQ ID NO: 423, respectively; h. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 51, SEQ ID NO: 142, and SEQ ID NO: 229, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 305, SEQ ID NO: 379, and SEQ ID NO: 429, respectively; i. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 60, SEQ ID NO: 152, and SEQ ID NO: 237, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 313, SEQ ID NO: 385, and SEQ ID NO: 435, respectively; j. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 64, SEQ ID NO: 157, and SEQ ID NO: 241, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 317, SEQ ID NO: 388, and SEQ ID NO: 438, respectively; k. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 69, SEQ ID NO: 161, and SEQ ID NO: 245, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 321, SEQ ID NO: 391, and SEQ ID NO: 441, respectively; l. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 72, SEQ ID NO: 166, and SEQ ID NO: 249, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 325, SEQ ID NO: 393, and SEQ ID NO: 444, respectively; m. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 77, SEQ ID NO: 171, and SEQ ID NO: 253, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 329, SEQ ID NO: 395, and SEQ ID NO: 447, respectively; n. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 82, SEQ ID NO: 172, and SEQ ID NO: 257, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 333, SEQ ID NO: 398, and SEQ ID NO: 450, respectively; or a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 100, SEQ ID NO: 166, and SEQ ID NO: 249, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 325, SEQ ID NO: 393, and SEQ ID NO: 444, respectively; p. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 105, SEQ ID NO: 191, and SEQ ID NO: 273, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 349, SEQ ID NO: 408, and SEQ ID NO: 462, respectively; q. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 110, SEQ ID NO: 196, and SEQ ID NO: 277, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 353, SEQ ID NO: 410, and SEQ ID NO: 465, respectively; r. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 115, SEQ ID NO: 201, and SEQ ID NO: 281, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 357, SEQ ID NO: 413, and SEQ ID NO: 468, respectively; s. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 120, SEQ ID NO: 205, and SEQ ID NO: 285, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 361, SEQ ID NO: 416, and SEQ ID NO: 471, respectively; t. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 488, SEQ ID NO: 497, and SEQ ID NO: 515, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 531, SEQ ID NO: 547, and SEQ ID NO: 558, respectively; or a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 489, SEQ ID NO: 502, and SEQ ID NO: 519, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 535, SEQ ID NO: 550, and SEQ ID NO: 561, respectively; v. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 489, SEQ ID NO: 506, and SEQ ID NO: 523, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 539, SEQ ID NO: 553, and SEQ ID NO: 564, respectively; or w. a VH region comprising a VH CDR1, a VH CDR2, and a VH CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 492, SEQ ID NO: 510, and SEQ ID NO: 527, respectively; and a VL region comprising a VL CDR1, a VL CDR2, and a VL CDR3 comprising the amino acid sequences set forth in SEQ ID NO: 543, SEQ ID NO: 556, and SEQ ID NO: 567, respectively.
  5. 5. Antibody or fragment thereof that specifically binds human GFRAL characterized in that the antibody or fragment binds: a. within amino acid residues 220-316 of SEQ ID NO: 1797; or b. to one or more residues selected from the group consisting of: Met214, Pro216, Pro217, Gln290, Cys291, Thr292, Cys293, Arg294, Thr295, Ile296, Thr297, Gln298, Ser299, Glu301, Lys305, Gln308, His309, His312 and Ser315 of SEQ ID NO: 1797.
  6. 6. Isolated polynucleotide characterized in that it encodes the antibody or a fragment thereof, as defined in any one of claims 1 to 5.
  7. 7. Vector characterized in that it comprises one or more polynucleotides encoding the antibody or a fragment thereof, as defined in any one of claims 1 to 5.
  8. 8. Pharmaceutical composition characterized in that it comprises the antibody or a fragment thereof, as defined in any one of claims 1 to 5, and a pharmaceutically acceptable carrier.
  9. 9. Use of the antibody or fragment thereof, as defined in any one of claims 1 to 5, characterized in that it is for the manufacture of a medicament to treat or prevent involuntary body weight loss in a human individual.
  10. 10. Method of producing the antibody or fragment thereof, as defined in any one of claims 1 to 5, characterized in that it comprises cultivating the cell comprising the isolated polynucleotide as defined in claim 6 or the vector as defined in claim 7, and purifying the antibody or fragment thereof.
  11. 11. Use of an antibody that specifically binds to a GDNF family receptor alpha-like protein (GFRAL) characterized in that it is for preparing a medicament for the treatment of cachexia or cancer cachexia, optionally wherein: (i) the antibody inhibits the binding of a human RET protein to a human GFRAL protein and interferes with an interaction between a human RET protein and the human GFRAL protein; and/or (ii) the antibody is administered at a dosage in the range of about 1 mg/kg to about 100.0 mg/kg every week or every three weeks, such that the subject receives from about two to about twenty doses, wherein said treatment increases the patient's body weight by at least about 5%; and/or (iii) the antibody has a dissociation constant (Kd) of less than 0.1 nM, measured by Biacore assay, by fluorescence-activated cell screening (FACS) assay or by an immunoassay, such as a radioimmunoassay (RIA) or immunoprecipitation; and/or (iv) the antibody inhibits human protein (GFRAL) activity and/or signaling with an IC50 value equal to or less than 10 nM in an ELK1 luciferase reporter assay or an ERK phosphorylation assay.

Description

FIELD [001] This application claims the benefit of Provisional Application in U.S. 62/316,516, filed March 31, 2016, the full contents of which are incorporated by reference. [002] The present disclosure relates generally to binding proteins, such as antibodies, that bind to a GDNF family receptor-type alpha protein (GFRAL), including human GFRAL protein, and methods for their use. BACKGROUND [003] Growth differentiation factor 15 (GDF15) is a protein belonging to the transforming growth factor beta (TGF-β) superfamily. GDF15 is also known as TGF-PL, MIC-1, PDF, PLAB, NAG-1, and PTGFB. GDF15 mRNA is reported to be most abundant in the liver, with lower levels seen in some other tissues. Its expression in the liver can be significantly upregulated in organ lesions such as liver, kidney, heart, and lung. [004] GDF15 is reported to play a role in regulating inflammatory and apoptotic pathways in injured tissues and during disease processes. GDF15 has been reported to be a mediator of cachexia in several diseases. However, cachexia is a complex and not fully understood syndrome. Additionally, at least some tumors overexpress and secrete GDF15, and elevated serum levels of GDF15 have been associated with several cancers. GDF-15 has been described as a negative regulator of macrophage activation by suppressing the release of TNF-α, IL-1, IL-2, and MCS-F, thus inhibiting positive feedback of local inflammatory signaling similar to the effects of TGF-β. Monoclonal antibodies against GDF15 have been revealed as potential therapeutic agents for the treatment of cachexia and cancer. The receptor for GDF15 is unknown. [005] There is a significant unmet need for effective therapeutic agents to treat weight loss associated with a variety of diseases and conditions, including wasting diseases such as cachexia or sarcopenia and inflammatory conditions such as systemic inflammation or an acute inflammatory response. There is also a significant unmet need for effective therapeutic agents to treat chronic diseases, including cancer, chronic kidney disease, chronic obstructive pulmonary disease, AIDS, tuberculosis, chronic inflammatory disease, systemic inflammation, and muscle wasting diseases, including those in which involuntary body weight loss and/or muscle mass loss is involved. SUMMARY [006] The present disclosure provides proteins that bind to a GDNF family receptor-type alpha protein (GFRAL), including binding proteins such as antibodies that bind to a GFRAL protein. Such binding proteins, including antibodies, may bind to a GFRAL polypeptide, a GFRAL fragment, and/or a GFRAL epitope. Such binding proteins, including antibodies, may be antagonistic (e.g., inhibit the binding of a GDF15 protein to a GFRAL protein, inhibit the binding of a RET protein to a GFRAL protein, inhibit GDF15-induced signaling, and/or inhibit the formation of a GDF15/GFRAL receptor complex or a GDF15/GFRAL/RET receptor complex). [007] The present disclosure also provides binding proteins, including antibodies (e.g., monoclonal antibodies) or fragments thereof, that (i) bind to a GFRAL protein, (ii) inhibit the binding of a GDF15 protein to a GFRAL protein, and/or (iii) inhibit the binding of a RET protein to a GFRAL protein. [008] In some embodiments, anti-GFRAL antibodies are humanized antibodies that bind to a GFRAL polypeptide, a GFRAL fragment, or a GFRAL epitope. In certain embodiments, an anti-GFRAL antibody comprises a VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 of a monoclonal antibody designated as 1C1, 3P10, 12A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2I23, 6N16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, or P8G4, as described herein, or a humanized variant thereof. In certain embodiments, an anti-GFRAL antibody may further comprise a VH FR1, VH FR2, VH FR3, VH FR4, VL FR1, VL FR2, VL FR3, and/or VL FR4 of an amino acid sequence of human immunoglobulin or a variant thereof. [009] In some embodiments, a binding protein (e.g., an anti-GFRAL antibody) comprises six or fewer CDRs. In some embodiments, a binding protein (e.g., an anti-GFRAL antibody) comprises one, two, three, four, five, or six CDRs selected from VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3. In some embodiments, a binding protein (e.g., an anti-GFRAL antibody) comprises one, two, three, four, five, or six CDRs selected from VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and/or VL CDR3 of a monoclonal antibody designated as 1C1, 3P10, 12A3, 5F12, 5A20, 8D8, 17J16, 25M22, 2B8, 22N5, 2I23, 6N16, 1B3, 19K19, 2B3, 8C10, 2A9, 24G2, 6G9, 2B11, 1A3, P1B6, P1H8, or P8G4, as described herein, or a humanized variant thereof. In some embodiments, a binding protein (e.g., an anti-GFRAL antibody) further comprises a framework region, including a VH FR1, VH FR2, VH FR3, VH FR4, VL FR1, VL FR2, VL FR3, and/or VL FR4 of an amino acid sequence of human immunoglobulin or a variant thereof. [010] In some embodiments, the antibody is a human