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EP-3555129-B1 - GREMLIN-1 CRYSTAL STRUCTURE AND INHIBITORY ANTIBODY

EP3555129B1EP 3555129 B1EP3555129 B1EP 3555129B1EP-3555129-B1

Inventors

  • DEDI, NEESHA
  • TWOMEY, Breda
  • WRIGHT, MICHAEL JOHN
  • DAVIES, GARETH
  • MCMILLAN, David James

Dates

Publication Date
20260506
Application Date
20171219

Claims (16)

  1. An anti-Gremlin-1 antibody which comprises a heavy chain variable region (HCVR) sequence comprising a HCDR1/HCDR2/HCDR3 sequence combination of SEQ ID NOs: 4/5/6 or SEQ ID NOs: 3/5/6 and a light chain variable region (LCVR) sequence comprising a LCDR1/LCDR2/LCDR3 sequence combination of SEQ ID NOs: 7/8/9.
  2. The antibody of claim 1, which comprises a HCVR sequence of SEQ ID NO: 10 or 12 and/or a LCVR sequence of SEQ ID NO: 11 or 13.
  3. The antibody of claim 2, which comprises a HCVR and LCVR sequence pair of SEQ ID NOs: 10/11 or 12/13.
  4. The antibody of claim 1, wherein the HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3 sequences consist of SEQ ID NOs: 4/5/6/7/8/9 or SEQ ID NOs: 3/5/6/7/8/9 and the remainder of the HCVR and LCVR sequences of the antibody comprise at least 95% identity to SEQ ID Nos: 10, 11, 12 and/or 13 respectively.
  5. The antibody of any one of claims 1-4, which comprises a heavy chain of SEQ ID NO: 14, 16, 18, 22, 28, 30, 32 or 34 and a light chain of SEQ ID NO: 15, 17, 19, 23, 29, 31, 33 or 35.
  6. The antibody of claim 5, which comprises a heavy and light chain pair of SEQ ID NOs: 14/15, 16/17, 18/19, 22/23, 28/29 or 30/31, 32/33, 34/35.
  7. The antibody of claim 1 or claim 4, wherein the HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3 sequences consist of SEQ ID NOs: 4/5/6/7/8/9 or SEQ ID NOs: 3/5/6/7/8/9 and the remainder of the heavy and light chains of the antibody comprise at least 95% identity to SEQ ID NOs: 14, 15, 16 and/or 17, respectively.
  8. An antibody according to claim 1, which is a chimeric, human or humanised antibody.
  9. An antibody according to any one of claims 1-8, which is a Fab, Fab', F(ab') 2 , Fv, or an scFv.
  10. An isolated polynucleotide encoding an antibody as defined in any one of claims 1-9.
  11. An expression vector carrying the polynucleotide of claim 10.
  12. A host cell comprising the vector as defined in claim 11.
  13. A method of producing the antibody as defined in any one of claims 1-9, comprising culturing the host cell of claim 12 under conditions permitting production of the antibody, and recovering the antibody produced.
  14. A pharmaceutical composition comprising an antibody as defined in any one of claims 1-9 and a pharmaceutically acceptable adjuvant and/or carrier.
  15. An antibody as defined in any one of claims 1-9 or a pharmaceutical composition as defined in claim 14 for use in a method of treatment of the human or animal body by therapy.
  16. The antibody or pharmaceutical composition for use according to claim 15, wherein renal fibrosis such as diabetic nephropathy, idiopathic pulmonary fibrosis, pulmonary arterial hypertension, angiogenesis or cancer are treated/prevented.

Description

Field of the Invention The invention provides antibodies which bind an allosteric inhibitory site on Gremlin-1, together with pharmaceutical compositions and medical uses of such antibodies. Background of the Invention Gremlin-1 (also known as Drm and CKTSF1B1) is a 184 amino acid glycoprotein which forms part of the DAN family of cystine-knot secreted proteins (along with Cerberus and Dan amongst others). Gremlin binds and inhibits the ability of BMP-2, 4, and 7 to signal along with a documented pro-angiogenic role possibly through agonism of VEGFR2. The main role of Gremlin-1 is during development, in which it is vital during kidney formation and during limb bud formation. These vital roles make gremlin homozygous knock-outs lethal in embryonic mice. In adulthood, increased levels of gremlin have been associated with idiopathic pulmonary fibrosis and pulmonary arterial hypertension in which BMP-2, 4 and 7 signalling is reduced with an associated rise in TGF-β levels. In both diabetic and chronic allograft nephropathy, Gremlin-1 expression has been correlated with fibrosis score. Increased levels of gremlin are also linked to scleroderma, diabetic nephropathy and colorectal cancer. Gremlin-1 has been shown to activate cancer cell invasion and proliferation and is thought to play a role in uterine cervix, lung, ovary, kidney, breast, colon, pancreatic and sarcoma carcinomas. To date, there have been a number of challenges associated with studying Gremlin-1 and there is a lack of general understanding around Gremlin-1 (and its partner Gremlin-2). BMP biology is complex, and high homology exists between species. Gremlin-1 is a difficult protein to work with, and there is a lack of suitable tools and reagents for studing its biology. Making Gremlin-1 is also not a straightforward process; cysteine-knot proteins are notoriously difficult to produce and the free cysteine of Gremlin-1 adds to the challenge. Gremlin-1 is difficult to express let alone purify. Until now, structural information has not been available and there is very little information on this protein in the literature. WO 2014/159010 A1 describes antibodies that bind to human gremlin-1 (GREM1) and methods of use. EP2826790 A1 describes a Gremlin-1 antibody that inhibits Gremlin-1 in a manner independent of bone morphogenetic protein (BMP) or vascular endothelial growth factor receptor-2 (VEGFR-2). Summary of the Invention The present invention, as defined by the appended claims, provides an anti-Gremlin-1 antibody which comprises a heavy chain variable region (HCVR) sequence comprising a HCDR1/HCDR2/HCDR3 sequence combination of SEQ ID NOs: 4/5/6 or SEQ ID NOs: 3/5/6 and a light chain variable region (LCVR) comprising a LCDR1/LCDR2/LCDR3 sequence combination of SEQ ID NOs: 7/8/9. The present invention also provides an isolated polynucleotide encoding the antibody of the invention. The present invention also provides an expression vector carrying the polynucleotide of the invention. The present invention also provides a host cell comprising the vector of the invention. The present invention also provides a method of producing the antibody of the invention, comprising culturing the host cell of the invention under conditions permitting production of the antibody, and recovering the produced antibody. The present invention also provides a pharmaceutical composition comprising an antibody of the invention and a pharmaceutically acceptable adjuvant and/or carrier. The present invention also provides an antibody of the invention or pharmaceutical composition of the invention for use in a method of treatment of the human or animal body by therapy. The term Gremlin-1 as used in the present invention typically has the sequence as set out in the UniProt entry O60565 (SEQ ID NO: 1). The term Gremlin-1 may also refer to a Gremlin-1 polypeptide which: (a) comprises or consists of the amino acid sequence of SEQ ID NO: 1 with or without the N-terminal signal peptide, i.e. may comprise or consist of the mature peptide sequence as shown in SEQ ID NO: 21; or(b) is a derivative having one or more amino acid substitutions, modifications, deletions or insertions relative to the amino acid sequence of SEQ ID NO: 1 with or without the N-terminal signal peptide (as shown in SEQ ID NO: 21), which retains the activity of Gremlin-1, such as the amino acid sequence of SEQ ID NO: 20.(c) a variant thereof, such variants typically retain at least about 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94% or 95% identity to SEQ ID NO: 1 (or SEQ ID NO: 20 or 21) (or even about 96%, 97%, 98% or 99% identity). In other words, such variants may retain about 60% - about 99% identity to SEQ ID NO: 1, suitably about 80% - about 99% identity to SEQ ID NO: 1, more suitably about 90% - about 99% identity to SEQ ID NO: 1 and most suitably about 95% - about 99% identity to SEQ ID NO: 1. Variants are described further below. As discussed further below, residue numbers are typically quoted based on the seq