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NZ766131B2 - Antibodies binding AXL - Google Patents
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NZ766131B2 - Antibodies binding AXL - Google Patents

Antibodies binding AXL Download PDF

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Publication number
NZ766131B2
NZ766131B2 NZ766131A NZ76613115A NZ766131B2 NZ 766131 B2 NZ766131 B2 NZ 766131B2 NZ 766131 A NZ766131 A NZ 766131A NZ 76613115 A NZ76613115 A NZ 76613115A NZ 766131 B2 NZ766131 B2 NZ 766131B2
Authority
NZ
New Zealand
Prior art keywords
antibody
region
seq
axl
cancer
Prior art date
Application number
NZ766131A
Other versions
NZ766131A (en
Inventor
Esther Breij
Jong Rob N De
Paul Parren
David Satijn
Den Brink Edward Norbert Van
Dijkhuizen Radersma Riemke Van
Dennis Verzijl
Original Assignee
Genmab A/S
Filing date
Publication date
Application filed by Genmab A/S filed Critical Genmab A/S
Priority claimed from NZ728019A external-priority patent/NZ728019A/en
Publication of NZ766131A publication Critical patent/NZ766131A/en
Publication of NZ766131B2 publication Critical patent/NZ766131B2/en

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    • A—HUMAN NECESSITIES
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    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3061—Blood cells
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/77—Internalization into the cell
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • G01N33/574—

Abstract

The present invention relates to anti-AXL antibodies, immunoconjugates, compositions and method of treatment of cancer with such anti-AXL antibodies, immunoconjugates, or compositions.

Claims (48)

Claims
1. An dy which binds to AXL, wherein the antibody does not compete for AXL binding with the ligand Growth Arrest-Specific 6 (Gas6) and wherein the antibody comprises at least one binding region, which comprises a le heavy chain (VH) region and a variable light chain (VL) region selected from the group consisting of: a) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 114, 115, and 116, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 117, DAS, and 118, tively [733]; b) a VH region comprising the CDR1, CDR2, and CDR3 ces of SEQ ID Nos.: 51, 52, and 53, respectively; and a VL region comprising the CDR1, CDR2, and CDR3 ces of SEQ ID Nos.: 55, GAS, and 56, respectively [154]; c) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 51, 52, and 54, respectively; and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 55, GAS, and 56, respectively [154-M103L]; d) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 72, 73, and 75, respectively; and a VL region sing the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 76, ATS, and 77, respectively, [183]; and e) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 72, 74, and 75, respectively; and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID Nos.: 76, ATS, and 77, respectively, [183-N52Q].
2. The antibody according to claim 1, wherein maximal antibody binding to AXL in the presence of Gas6 is at least 90%, such as at least 95%, such as at least 97%, such as at least 99%, such as 100%, of binding in the e of Gas6 as determined by a competition assay, wherein competition between said antibody binding to AXL and said Gas6 is determined on A431 cells preincubated with Gas6 and without Gas6.
3. The antibody according to any one of claims 1 or 2, n said antibody has a binding affinity (KD) in the range of 0.3x10-9 to 63x10-9 M to human AXL, and wherein said binding affinity is measured using a Bio-layer Interferometry using soluble AXL extracellular domain. 20631521_1 (GHMatters) P42488NZ02
4. The antibody according to any one of the preceding claims, wherein said antibody has a dissociation rate of 9.7x10-5 to 4.4x10-3 s-1 to AXL, and wherein the dissociation rate is measured by Bio-layer Interferometry using soluble recombinant AXL extracellular domain.
5. The antibody according to any one of the preceding claims, wherein AXL is human AXL as specified in SEQ ID NO:130.
6. The dy according to any one of claims 1 to 4, wherein AXL is cynomolgus monkey AXL as specified in SEQ ID NO:147.
7. The antibody ing to any one of the preceding claims, wherein AXL is human AXL as specified in SEQ ID NO:130 and cynomologus monkey AXL as specied in SEQ ID NO:147.
8. The antibody according to any one of the preceding claims, wherein said at least one binding region comprises a VH region and a VL region selected from the group consisting of; a) a VH region comprising SEQ ID No: 34 and a VL region sing SEQ ID No: 35 [733]; b) a VH region comprising SEQ ID No: 7 and a VL region comprising SEQ ID No: 9 [154]; c) a VH region comprising SEQ ID No: 16 and a VL region sing SEQ ID No: 18 [183]; d) a VH region comprising SEQ ID No:8 and a VL region comprising SEQ ID No:9 [154- M103L]; and e) a VH region comprising SEQ ID No:17 and a VL region comprising SEQ ID No:18 [183- N52Q].
9. The antibody according to any one of claims 1 to 7, wherein said antibody binds to an epitope on AXL wherein said epitope is recognized by any of the antibodies defined in claim 8.
10. The dy according to any one of the preceding claims, n said antibody comprises a heavy chain of an isotype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4.
11. The antibody according to claim 10, n the isotype is IgG1, optionally pe IgG1m(f).
12. The antibody of any one of the preceding claims, which is a full-length onal antibody, such as a full-length monoclonal IgG1,? antibody. 20864917_1 (GHMatters) P42488NZ02
13. The antibody according to any one of claims 1 to 12, wherein said antibody is an effector-function- deficient dy, a stabilized IgG4 dy or a monovalent antibody.
14. The antibody according to claim 13, n said heavy chain has been modified such that the entire hinge region has been deleted.
15. The dy according to any one of claims 12 and 13, wherein the sequence of said antibody has been modified so that it does not comprise any acceptor sites for N-linked glycosylation.
16. The antibody according to any one of claims 1 to 15, wherein said antibody is a single-chain antibody.
17. A bispecific antibody sing a first binding region of an antibody according to any one of the preceding claims, and a second binding region which binds a different target or epitope than said first binding region.
18. The bispecific antibody according to claim 17, n said bispecific antibody comprises a first and a second heavy chain, each of said first and second heavy chain comprises at least a hinge region, a CH2 and CH3 region, wherein in said first heavy chain at least one of the amino acids in the positions corresponding to positions selected from the group consisting of K409, T366, L368, K370, D399, F405, and Y407 in a human IgG1 heavy chain has been substituted, and in said second heavy chain at least one of the amino acids in the positions corresponding to a position selected from the group consisting of F405, T366, L368, K370, D399, Y407, and K409 in a human IgG1 heavy chain has been substituted, and wherein said tutions of said first and said second heavy chains are not in the same positions.
19. The bispecific antibody according to claim 18, wherein the amino acid in the on corresponding to K409 in a human IgG1 heavy chain is R in said first heavy chain, and the amino acid in the position corresponding to F405 in a human IgG1 heavy chain is L in said second heavy chain, or vice versa.
20. An immunoconjugate comprising the antibody ing to any one of claims 1 to 16, or bispecific antibody according to any one of claims 17 to 19, and a therapeutic moiety, such as a cytotoxic agent, a chemotherapeutic drug, a cytokine, an immunosuppressant, antibiotic, or a radioisotope. 20864917_1 (GHMatters) P42488NZ02
21. The immunoconjugate ing to claim 20, n the therapeutic moiety is a cytotoxic agent.
22. The immunoconjugate according to claim 20 or 21, wherein said cytotoxic agent is linked to said antibody with a linker.
23. The immunoconjugate according to any one of claims 20 to 22, wherein said cytotoxic agent is linked to said antibody with a cleavable linker, such as N-succinimydyl yridyldithio)-pentanoate (SSP), maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-vc-PAB) or AV-1 K-lock valine- citrulline.
24. The immunoconjugate ing to any one of claims 20 to 22, wherein said cytotoxic agent is linked to said antibody, or fragment f, with a non-cleavable linker, such as succinimidyl-4(N- maleimidomethyl)cyclohexanecarboxylate (MCC) or maleimidocaproyl (MC).
25. The immunoconjugate ing to any one of claims 20 to 24, wherein said cytotoxic agent is selected from the group: DNA-targeting agents, e.g. DNA alkylators and cross-linkers, such as calicheamicin, duocarmycin, rachelmycin (CC-1065), pyrrolo[2,1-c][1,4] benzodiazepines (PBDs), and indolinobenzodiazepine (IGN); microtubule-targeting agents, such as duostatin, such as duostatin-3, auristatin, such as monomethylauristatin E (MMAE) and monomethylauristatin F (MMAF), dolastatin, maytansine, N(2’)-deacetyl-N(2’)-(3-marcaptooxopropyl)-maytansine (DM1), and sin; and nucleoside analogs; or an analogs, derivatives, or prodrugs thereof.
26. The immunoconjugate according to any one of the claims 22 to 25, wherein said conjugate comprises a combination of; i) said cytotoxic agent and said cleavable linker having bystander kill capacity; ii) said cytotoxic agent and said ble linker not having bystander kill capacity; iii) said cytotoxic agent and said non-cleavable linker having bystander kill capacity; or iv) said xic agent and said non-cleavable linker not having bystander kill capacity.
27. The immunoconjugate according to any one of claims 22 to 23, and 25 to 26, wherein said linker is mc-vc-PAB and the cytotoxic agent is MMAE; or the linker is SSP and the cytotoxic agent is DM1. 20864917_1 (GHMatters) P42488NZ02
28. The conjugate according to any one of the claims 22 to 23 and 25 to 26, wherein said conjugate comprises said linker mc-vc-PAB, said cytotoxic agent MMAE and said antibody wherein said at least one binding region comprises a VH region and a VL region selected from the group consisting of; a) a VH region comprising SEQ ID No: 34 and a VL region comprising SEQ ID No: 35 [733] b) a VH region comprising SEQ ID No: 7 and a VL region comprising SEQ ID No: 9 [154]; c) a VH region comprising SEQ ID No: 16 and a VL region comprising SEQ ID No: 18 [183]; d) a VH region comprising SEQ ID No:8 and a VL region comprising SEQ ID No:9 [154- M103L]; and e) a VH region comprising SEQ ID No:17 and a VL region comprising SEQ ID No:18 [183- N52Q];
29. The immunoconjugate according to any one of claims 22, and 24 to 25, wherein said linker is MMC and the cytotoxic agent is DM1; or the linker is MC and the cytotoxic agent is MMAF.
30. The immunoconjugate according to any one of claims 20 to 29, wherein the number of cytotoxic agents per antibody is from 1 to 8, such as 2 to 7, such as 2 to 6, such as 2 to 5, such as 2 to 4, and such as 2 to 3.
31. A composition comprising the antibody ing to any one of claims 1 to 16, a bispecific antibody according to any one of claims 17 to 19, or an immunoconjugate according to any one of claims 20 to
32. A pharmaceutical composition comprising the antibody ing to any one of claims 1 to 16, a bispecific dy according to any one of claims 17 to 19, or an immunoconjugate according to any one of claims 20 to 30, and a pharmaceutical acceptable carrier.
33. A nucleic acid construct encoding an antibody according to any one of claims 1 to 16.
34. An expression vector comprising one or more nucleic acid ucts ing to claim 33.
35. An isolated and/or non-human host cell comprising a vector according to claim 34. 20864917_1 (GHMatters) P42488NZ02
36. The host cell according to claim 35, wherein said host cell is a recombinant host cell, such as a recombinant prokaryotic, recombinant eukaryotic, or recombinant microbial host cell.
37. The host cell according to any one of claims 35 to 36, wherein the host cell produces an antibody ing to any one of claims 1 to 16, or a bispecific antibody according to any one of claims 17 to 19.
38. Use of the antibody ing to any one of claims 1 to 16, the bispecific antibody according to any one of claims 17 to 19, or the immunoconjugate according to any one of claims 20 to 30, the ition according to claim 31, or the pharmaceutical composition according to claim 32, in the manufacture of a medicament for treating cancer.
39. A method of diagnosing a e characterized by involvement or accumulation of AXL-expressing cells, wherein an antibody according to any one of claims 1 to 16, a bispecific antibody according to any one of claims 17 to 19, an immunoconjugate according to any one of claims 20 to 30, a ition according to claim 31, or a pharmaceutical composition ing to claim 32, is contacted with a sample from a subject, optionally wherein the antibody is labeled with a detectable agent, and wherein the amount of AXL-expressing cells correlates with or is indicative of disease.
40. The method according to claim 39, wherein the disease is cancer.
41. The use according to claim 38, or the method according to claim 40, wherein the cancer is a solid tumor sing AXL or an AXL-expressing hematological cancer.
42. The use or method according to claim 41, wherein the hematological cancer is selected from the group consisting of leukemia, such as chronic cytic leukemia, myeloid ia, acute d leukemia (AML) and chronic myeloid ia, lymphoma such as Non-Hodgkin lymphoma and multiple myeloma.
43. The use or method according to claim 41, wherein the solid tumor expressing AXL is lung cancer or epidermoid carcinoma.
44. The use or method according to any one of claims 38 and 40, wherein the cancer is selected from the group consisting of colorectal cancer, such as colorectal carcinoma and colorectal adenocarcinoma, 20864917_1 (GHMatters) P42488NZ02 bladder cancer, bone cancer such as chondrosarcoma, breast cancer such as triple-negative breast cancer, cancers of the central nervous system such as astoma, astrocytoma, neuroblastoma, cervical cancer, connective tissue cancer, endometrium cancer, fibroblast cancer, gastric cancer such as gastric carcinoma, head and neck cancer, kidney , liver cancer such as hepatocellular carcinoma, lung cancer such as NSCLC and lung squamous cell carcinoma, muscle cancer, neural tissue cancer, ovarian cancer, atic cancer such as pancreatic ductal carcinoma and pancreatic arcinoma, skin cancer such as malignant melanoma and soft tissue sarcoma.
45. Use of an antibody ing to any one of claim 1 to 16, a bispecific antibody according to any one of claims 17 to 19, an immunoconjugate according to any one of claims 20 to 30, a composition according to claim 31, or a pharmaceutical composition according to claim 32, in the manufacture of a medicament for inhibiting growth and/or proliferation of a tumor cell expressing AXL.
46. A method for producing an antibody of any one of claims 1 to 16, said method comprising the steps a) culturing a host cell according to any one of claims 35 to 37, and b) purifying the antibody from the culture media.
47. A method for detecting the presence of AXL dy, or a cell expressing AXL, in a sample comprising the steps of; a) contacting the sample with an antibody according to any one of claims 1 to 16, a bispecific antibody according to any one of claims 17 to 19, or an conjugate according to any one of claims 20 to 30, under conditions that allow for formation of a complex between the dy, bispecific antibody or immunoconjugate and AXL; and b) analyzing whether a complex has been formed.
48. A kit for detecting the presence of AXL antigen, or a cell expressing AXL, in a sample comprising i) an antibody ing to any one of claims 1 to 16, a bispecific antibody according to any one of claims 17 to 19, or an immunoconjugate according to any one of claims 20 to 30; and ii) instructions for use of said kit. 20864917_1 (GHMatters) P42488NZ02 1/ 25
NZ766131A 2015-07-10 Antibodies binding AXL NZ766131B2 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
DKPA201400380 2014-07-11
DKPA201400489 2014-09-01
DKPA201400746 2014-12-22
DKPA201500283 2015-05-12
NZ728019A NZ728019A (en) 2014-07-11 2015-07-10 Antibodies binding axl

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NZ766131B2 true NZ766131B2 (en) 2024-10-08

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