Anti-gpvi antibodies and functional fragments thereof

Abstract

The present invention relates to antibody molecules and functional fragments thereof, capable of binding to human glycoprotein VI (GPVI), to processes for their production, and to their therapeutic uses

Description

Abstract This invention relates to antibody molecules capable of binding to human glycoprotein VI (GPVI) and their functional fragments, methods of their production, and their therapeutic uses.

Claims

Claims1. An antibody or a functional fragment thereof capable of binding to human glycoprotein VI (GPVI), wherein the antibody or functional fragment comprises (i) a VLdomain comprising a CDR1 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:1 , a CDR2 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:2 or SEQ ID NO:9 or SEQ ID NO:64, and a CDR3 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:3, and (ii) a H domain comprising a CDR1 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:4, a CDR2 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:5, and a CDR3 region having an amino acid sequence in accordance with the amino acid sequence as shown in SEQ ID NO:6.

2. The antibody or functional fragment of claim 1 , which is monovalent.

3. The antibody or functional fragment of claim 1 or 2, wherein the antibody or functional fragment binds to human GPVI with a dissociation equilibrium constant (KD) of less than 700 pM, preferably less than 300 pM, more preferably less than 200 pM.

4. The antibody or functional fragment of any one of the preceding claims, wherein the antibody or functional fragment comprises a VL domain comprising an amino acid sequence having a sequence identity of at least 90 % to a sequence selected from the group consisting of SEQ ID NO:16, 17, 18, 19, 26, 27, 28, 29, 30, 31 , 32, 33, 34, and 35; and / or a VHdomain comprising an amino acid sequence having a sequence identity of at least 90 % to a sequence selected from the group consisting of SEQ ID NO:21 , 22, 23, 24 and 25.

5. The antibody or functional fragment of any one of the preceding claims, wherein the antibody or functional fragment comprises a VL domain comprising an amino acid sequence as shown in SEQ ID NO:16, 17, 18, 19, 26, 27, 28, 29, 30, 31 , 32, 33, 34, or 35; and / or a VH domain comprising an amino acid sequence as shown in SEQ ID NO:21 , 22, 23, 24 or 25.

6. The functional fragment of any one of the preceding claims, which is a fragment antigen-binding (Fab), a F(ab’), an Fv, a monovalent IgG, a disulfide-linked variable fragment (dsFv), or a single-chain variable fragment (scFv).

7. The antibody or functional fragment of any one of the preceding claims, wherein said antibody or functional fragment comprises a light chain comprising an N-terminal light chain signal peptide having an amino acid sequence in accordance with the sequence as shown in SEQ ID NO:62 and / or a constant domain having an amino acid sequence as shown in SEQ ID NO:60; and / or a heavy chain comprising an N-terminal light chain signal peptide having an amino acid sequence in accordance with the sequence as shown in SEQ ID NO:63 and / or a constant domain having an amino acid sequence as shown in SEQ ID NO:61.

8. The functional fragment of claim 7, wherein the functional fragment is a Fab, comprising, or consisting of, a light chain having an amino acid sequence as shown in one of SEQ ID NO: 38, 39, 40, 41 , 55, 56, 57 or 58, and a heavy chain having an amino acid sequence as shown in one of SEQ ID NO:44, 45, 46, 47 or 48.

9. The functional fragment of any one of the preceding claims, wherein the functional fragment is a Fab consisting of a pair of light chain and heavy chain, having the amino acid sequence as shown SEQ ID NO:39 and 44, 39 and 45, 40 and 45, 56 and 44, 56 and 45, or 57 and 45, preferably SEQ ID NO:39 and 44, or 56 and 44.

10. The antibody or functional fragment of any one of the preceding claims, which is capable of binding human GPVI at a binding epitope corresponding to amino acids V178 to E192 and / or S223 to P237 of SEQ ID NO:36.

11. A nucleic acid encoding the antibody or functional fragment of any one of the preceding claims.

12. A cell comprising the nucleic acid of claim 11 .

13. A method of preparing the antibody or functional fragment of any one of claims 1 to 10, comprising culturing the cell of claim 12 in a medium under conditions that allow expression of the nucleic acid encoding the antibody or functional fragment, and recovering the antibody or functional fragment from the cells or from the medium.

14. A pharmaceutical composition comprising the antibody or functional fragment of any one of claims 1 to 10, and optionally a pharmaceutically acceptable carrier and / or excipient.

15. The antibody or functional fragment as defined in any one of claims 1 to 10, or the pharmaceutical composition of claim 14, for use in a method of treating or preventing a GPVI-related condition in a subject; optionally wherein the GPVI-related condition is a thrombo-inflammatory disease; a cardiovascular disease, preferably selected from thrombosis and thrombotic disorders, including arterial thrombosis, venous thrombosis, atherothrombosis, stent thrombosis, venous thromboembolism diseases, thrombotic microangiopathies, cancer-associated thrombosis, immunothrombosis and thrombosis associated to infection, restenosis, acute coronary syndrome, ischemic cerebrovascular accidents, cerebral vascular diseases, vascular purpura, coronary artery diseases and cerebral artery diseases, ischemic events, acute coronary artery syndrome, myocardial infarction, stroke, percutaneous coronary intervention, ischemic restenosis, acute ischemia, chronic ischemia, diseases of the aorta and its branches, peripheral artery disease, acute phlebitis, pulmonary embolism; inflammation, preferably sustained or prolonged inflammation associated with infection, arthritis, fibrosis, acute respiratory distress syndrome (ARDS), ischemia-reperfusion injury (IRI) of various organs (liver, colon, etc.), peripheral vascular disease, antiphospholipid syndrome (APS), deep vein thrombosis, thrombophlebitis & vasculitis, transfusion- related acute lung injury (TRALI), transplant rejection, pre-eclampsia, severe burns, atherosclerosis, hypertension, antiphospholipid syndrome, preeclampsia, sickle cell disease, bacterial and viral infection ischemic restenosis, sepsis, major trauma, autoimmune diseases, and disorders in which platelets modulate cell functions including, without limitation, cancer cells proliferation and / or dissemination; cancer, preferably skin cancer, colon cancer, breast cancer, ovarian cancer, lung cancer, or metastatic cancer; and / or a disorder associated with abnormal or aberrant megakaryocyte and / or platelet proliferation, differentiation, morphology, migration, aggregation, degranulation and / or function.