LOX and LOXL2 inhibitors and uses thereof

• US 10,494,443 B2
• Filed: 11/02/2016
• Issued: 12/03/2019
• Est. Priority Date: 08/02/2007
• Status: Active Grant

• Current Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Original Assignee: Gilead Biologics, Inc. (Gilead Sciences Inc.)
• Inventors: Smith, Victoria, Ogg, Scott, Van Vlasselaer, Peter, Barry, Vivian E., Marshall, Derek, Holzer, Alison Kay, Rodriguez, Hector, Oyasu, Miho, McCauley, Scott Alan, Garcia, Carlos Aurelio, Biermann, Donna Hiroko Tokuoka
• Primary Examiner(s): Sang, Hong
• Application Number:
  US15/341,729
• Publication Number:
  US 20170240649A1
• Time in Patent Office: 1,126 Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 13/10 of the bladder
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 : specific for metastasis
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: C07K 16/40 : against enzymes
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/30 : characterized by aspects of specificity or valency
• CPC Class Codes: C07K 2317/33 : Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/34 : Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/77 : Internalization into the cell
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12Q 1/6886 : for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: G01N 2333/90633 : with oxygen as acceptor (1.4.3) in general
• CPC Class Codes: G01N 33/57423 : of lung
• CPC Class Codes: G01N 33/57438 : of liver, pancreas or kidney
• CPC Class Codes: G01N 33/57484 : involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites

Abstract

The present application relates to anti-LOX and anti-LOXL2 antibodies and their use in purification, diagnostic and therapeutic methods. Antibodies include monoclonal antibodies, humanized antibodies and functional fragments thereof. Anti-LOX and anti-LOXL2 antibodies can be used to identify and treat conditions such as a fibrotic condition, angiogenesis, or to prevent a transition from an epithelial cell state to a mesenchymal cell state.

Claims

1. An isolated monoclonal antibody or antigen binding fragment thereof that specifically binds to a lysyl oxidase-like 2 (LOXL2) protein and comprises a heavy chain variable region comprising complementarity determining region (CDR)1, CDR2 and CDR3 comprising the amino acid sequences set forth as SEQ ID NOs:

41, 42, and 70, respectively, and a light chain variable region comprising CDR1, CDR2 and CDR3 comprising the amino acid sequences set forth as SEQ ID NOs;

57, 58, and 59, respectively.

2. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment inhibits the enzymatic activity of the LOXL2 protein.

3. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment is a non-competitive inhibitor of the LOXL2 protein.

4. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment inhibits LOXL2 binding to ECM proteins, cellular receptors, or integrins.

5. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment reduces or inhibits uptake or internalization of LOXL2 protein.

6. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment is humanized.

7. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment is labeled with a therapeutic agent.

8. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment is labeled with a detectable label.

9. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment binds to LOXL2 with a binding affinity of at least 2, 5, 10, 50, 100, 500, or 1000 times greater than the binding affinity to at least one of LOX, LOXL1, LOXL3, or LOX4 proteins.

10. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment inhibits LOXL2 biological activity.

11. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment reduces tumor growth.

12. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the antibody or antigen binding fragment reduces incidence of metastasis.

13. A pharmaceutical composition comprising the monoclonal antibody or antigen binding fragment of claim 1 and a pharmaceutically acceptable excipient.

14. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the heavy chain variable region comprises the amino acid sequence set forth as SEQ ID NOs:

25, 26, 27, or 28.

15. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the heavy chain variable region comprises the amino acid sequence set forth as SEQ ID NO:

27.

16. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the light chain variable region comprises the amino acid sequence set forth as SEQ ID NOs:

30, 31, or 32.

17. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the light chain variable region comprises the amino acid sequence set forth as SEQ ID NO:

31.

18. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the heavy chain variable region comprises the amino acid sequence set forth as SEQ ID NO:

27 and the light chain variable region comprises the amino acid sequence set forth as SEQ ID NO;

31.

19. The isolated monoclonal antibody or antigen binding fragment of claim 1, wherein the isolated antibody or antigen binding fragment thereof is a Fv, a scFv, a Fab or a F(ab′

)₂.

20. The pharmaceutical composition of claim 13, further comprising a second therapeutic agent.

21. The pharmaceutical composition of claim 20, wherein the second therapeutic agent is an antibody or an anti-fibrotic agent.

22. The pharmaceutical composition of claim 21, wherein the anti-fibrotic agent is selected from the group consisting of ethylenediamine, hydrazine, phenylhydrazine, semicarbazide, urea, aminonitriles, beta-aminopropionitrile (BAPN), 2-nitroethylamine, unsaturated or saturated haloamines, 2-bromo-ethylamine, 2-chloroethylamine, 2-trifluoroethyl amine, 3-bromopropylamine, p-halobenzylamines, selenohomocysteine lactone, copper chelating agents, thiolamines, D-penicillamine, 2-amino-5-mercapto-5-methylhexanoic acid, D-2-amino-3-methyl-3-((2-acetamidoethyl)dithio)butanoic acid, p-2-amino-3-methyl-3-((2-aminoethyl)dithio)butanoic acid, sodium-4-((p-1-dimethyl-2-amino-2-carboxyethyl)dithio) butane sulphinate, 2-acetamidoethyl-2-acetamidoethanethiol sulphanate, and sodium-4-mercaptobutanesulphinate trihydrate.