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WO 2014/152006 A2 25 September 2014 (25.09.2014) P O P C T

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WO 2014/152006 A2 25 September 2014 (25.09.2014) P O P C T (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2014/152006 A2 25 September 2014 (25.09.2014) P O P C T (51) International Patent Classification: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, A61K 39/395 (2006.01) BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (21) International Application Number: HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, PCT/US20 14/026804 KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, (22) International Filing Date: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 13 March 2014 (13.03.2014) OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, (25) Filing Language: English TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, (26) Publication Language: English ZW. (30) Priority Data: (84) Designated States (unless otherwise indicated, for every 61/791,953 15 March 2013 (15.03.2013) US kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, (71) Applicant: INTRINSIC LIFESCIENCES, LLC UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, [US/US]; 505 Coast Boulevard South, Suite 408, La Jolla, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, California 92037 (US). EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, (72) Inventors: WESTERMAN, Mark; 5094 Mt. La Platta TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Dr., San Diego, California 921 17 (US). OSTLAND, KM, ML, MR, NE, SN, TD, TG). Vaughn; 79449 Calle Vista Verde, La Quinta, California 92253 (US). HAN, Huiling; 12396 Picrus Street, San Declarations under Rule 4.17 : Diego, California 9212 (US). GUTSCHOW, Patrick; 2 11 — as to applicant's entitlement to applyfor and be granted a Rainbow Lane, Oceanside, California 92054 (US). WEST¬ patent (Rule 4.1 7(H)) ERMAN, Keith; 5094 Mt. La Platta Drive, San Diego California 921 17 (US). OLBINA, Gordana; 709 Silver Published: Street, La Jolla, California 92037 (US). — without international search report and to be republished (74) Agents: GUISE, Jeffrey W. et al; WILSON SONSINI upon receipt of that report (Rule 48.2(g)) GOODRICH & ROSATI, 650 Page Mill Road, Palo Alto, — with sequence listingpart of description (Rule 5.2(a)) California 94304 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, (54) Title: ANTI-HEPCIDIN ANTIBODIES AND USES THEREOF FIGURE 2.0- • Hepcidin-25 Hepcidin-22 1.5 Hepcidin-20 < 0.5- © 0.0- o - 4 -2 0 2 Hepcidin (ng) (57) Abstract: The present application relates to antibodies that specifically bind to hepcidin and methods of using the antibodies. Another aspect relates to antibodies which bind hepcidin and regulate iron homeostasis. Another aspect relates to the use of human - ized antibodies which bind hepcidin for the treatment of a disease or condition associated with hepcidin. ANTI-HEPCIDIN ANTIBODIES AND USES THEREOF CROSS REFERENCE [0001] This application claims the benefit of U.S. Provisional Application No. 61/791,953, filed March 15, 2013, which application is incorporated herein by reference in its entirety. SEQUENCE LISTING [0002] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on March 7, 2014, is named 44546-702.601_SL.txt and is 38,025 bytes in size. BACKGROUND OF THE INVENTION [0003] Iron is an essential trace element required for growth and development of living organisms. In mammals, iron content is regulated by controlling iron absorption, iron recycling, and release of iron from the cells in which it is stored. Iron is predominantly absorbed in the duodenum and upper jejunum by enterocytes. Iron is recycled from degraded red cells by reticuloendothelial macrophages in bone marrow, hepatic Kupffer cells and spleen. Iron release is controlled by ferroportin, a major iron export protein located on the cell surface of enterocytes, macrophages and hepatocytes, the main cells capable of releasing iron into plasma. Hepcidin binds to ferroportin and decreases its functional activity by causing it to be internalized from the cell surface and degraded. (Nemeth et a , Science, 306:2090-3, 2004). SUMMARY OF THE INVENTION [0004] Provided herein are antibodies and antigen-binding fragments thereof that bind to hepcidin (Hep) or a hepcidin peptide. In one aspect, provided herein is an antibody, or antigen- binding fragment thereof, that specifically binds to the N-terminus of hepcidin or a hepcidin peptide and neutralizes the activity of hepcidin in vitro and/or in vivo. [0005] In one aspect, provided herein is an antibody, or antigen-binding fragment thereof, that specifically binds to hepcidin or a hepcidin peptide, comprising a heavy chain variable region and a light chain variable region, [0006] wherein said heavy chain variable region comprises: [0007] (i) a CDR1 having an amino acid sequence of any one of SEQ ID NOS: 55-57, [0008] (ii) a CDR2 having an amino acid sequence of any one of SEQ ID NOS: 58-60, and [0009] (iii) a CDR3 having an amino acid sequence of any one of SEQ ID NOS: 61-63; [0010] and said light chain variable region comprises: [0011] (i) a CDR1 having an amino acid sequence of any one of SEQ ID NOS: 64-66, [0012] (ii) a CDR2 having an amino acid sequence of any one of SEQ ID NOS: 67-69, and [0013] (iii) a CDR3 having an amino acid sequence of any one of SEQ ID NOS: 70-72. [0014] In one aspect, provided herein is an antibody, or antigen-binding fragment thereof, that specifically binds to hepcidin or a hepcidin peptide, comprising a heavy chain variable region and a light chain variable region, [0015] wherein said heavy chain variable region comprises: [0016] (i) a CDR1 having an amino acid sequence encoded by any one of SEQ ID NOS: 1-3, [0017] (ii) a CDR2 having an amino acid sequence encoded by any one of SEQ ID NOS: 4-6, and [0018] (iii) a CDR3 having an amino acid sequence encoded by any one of SEQ ID NOS: 7-9; [0019] and said light chain variable region comprises: [0020] (i) a CDR1 having an amino acid sequence encoded by any one of SEQ ID NOS: 10-12, [0021] (ii) a CDR2 having an amino acid sequence encoded by any one of SEQ ID NOS: 13-15, and [0022] (iii) a CDR3 having an amino acid sequence encoded by any one of SEQ ID NOS: 16- 18. [0023] In one aspect, an antibody, or antigen-binding fragment thereof, provided herein comprises IgGl or an IgG4 variable heavy chain and variable light chain. [0024] In one aspect, provided herein is an antibody, or antigen-binding fragment thereof, that specifically binds to hepcidin or a hepcidin peptide, that is prepared by injecting a rodent (i.e., mouse, rat or rabbit) with a peptide having an amino acid sequence of any one of SEQ ID NOS: 19-27. In another embodiment, the peptide is conjugated to a carrier (e.g., keyhole limpet hemocyanin (KLH)) or administered with an adjuvant (complete Freund's adjuvant (CFA) or incomplete Freund's adjuvant (IFA)). In another embodiment the peptide is conjugated to a hapten (e.g., dinitrophenol [DNP]) and a carrier. [0025] A hepcidin peptide to which an antibody, or antigen-binding fragment thereof, binds may have, in some instances, an amino acid sequence of SEQ ID NO: 19. [0026] Provided herein is an antibody, or antigen-binding fragment thereof, that specifically binds to an epitope comprising amino acid sequence of any one of Hep-5, Hep-9, Hep-20, Hep 22 and Hep25. [0027] In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds to an epitope comprising an amino acid sequence of Hep-20 (SEQ ID NO: 22), Hep 22 (SEQ ID NO: 23) and Hep25 (SEQ ID NO: 19). [0028] In one embodiment, the antibody, or antigen-binding fragment thereof, specifically binds to an epitope comprising Hep-5 (SEQ ID NO: 25) or Hep-9 (SEQ ID NO: 24). In another embodiment, provided herein is an antibody, or antigen-binding fragment thereof, that specifically binds to an epitope comprising amino acid residues 1-9 of hepcidin. In another embodiment, the antibody, or antigen-binding fragment thereof, specifically binds to 2, 3, 4, 5, 6, 7, 8, or 9 amino acid residues of an epitope comprising amino acid residues 1-9 of hepcidin. [0029] In another embodiment, the antibody, or antigen-binding fragment thereof, is monoclonal antibody comprising a heavy chain CDR1 encoded by SEQ ID NO: 55, a heavy CDR2 encoded by SEQ ID NO: 58, a heavy chain CDR3 encoded by SEQ ID NO: 61, a light chain CDR1 encoded by SEQ ID NO: 64, a light CDR2 encoded by SEQ ID NO: 67, and a light chain CDR3 encoded by SEQ ID NO: 70. [0030] In another embodiment, the antibody, or antigen-binding fragment thereof, is monoclonal antibody comprising a heavy chain CDR1 encoded by SEQ ID NO: 56, a heavy CDR2 encoded by SEQ ID NO: 59, a heavy chain CDR3 encoded by SEQ ID NO: 61, a light chain CDR1 encoded by SEQ ID NO: 65, a light CDR2 encoded by SEQ ID NO: 68, and a light chain CDR3 encoded by SEQ ID NO: 71.
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