Peptide core-based multi-arm linkers

• US 9,907,858 B2
• Filed: 01/18/2016
• Issued: 03/06/2018
• Est. Priority Date: 01/16/2015
• Status: Active Grant

• Current Assignee: Immunwork Inc.
• Original Assignee: Immunwork Inc.
• Inventors: Chang, Tse-Wen, Chu, Hsing-Mao
• Primary Examiner(s): Canella, Karen A.
• Application Number:
  US14/997,764
• Publication Number:
  US 20160208020A1
• Time in Patent Office: 778 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 31/397 having four-membered rings, e.g. azetidine
• CPC Class Codes: A61K 31/4545 containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
• CPC Class Codes: A61K 31/4709 Non-condensed quinolines and containing further heterocyclic rings
• CPC Class Codes: A61K 31/4745 condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines yohimbine derivatives, vinblastine A61K31/475; ergoline derivatives A61K31/48
• CPC Class Codes: A61K 31/537 spiro-condensed or forming part of bridged ring systems
• CPC Class Codes: A61K 31/739 Lipopolysaccharides
• CPC Class Codes: A61K 47/58 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/61 the organic macromolecular compound being a polysaccharide or a derivative thereof
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/6801 Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
• CPC Class Codes: A61K 47/6803 Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
• CPC Class Codes: A61K 47/6843 the antibody targeting a material from animals or humans
• CPC Class Codes: A61K 47/6845 the antibody targeting a cytokine, e.g. growth factors, VEGF, TNF, a lymphokine or an interferon
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: A61K 47/6883 Polymer-drug antibody conjugates, e.g. mitomycin-dextran-Ab; DNA-polylysine-antibody complex or conjugate used for therapy
• CPC Class Codes: A61K 51/065 conjugates with carriers being macromolecules
• CPC Class Codes: A61K 51/088 conjugates with carriers being peptides, polyamino acids or proteins antibodies A61K51/10
• 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 17/06 : Antipsoriatics
• CPC Class Codes: A61P 19/02 : for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/10 : for osteoporosis
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 35/02 : specific for leukemia
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 : Immunomodulators
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 14/485 : Epidermal growth factor [EGF], i.e. urogastrone
• CPC Class Codes: C07K 14/655 : Somatostatins
• CPC Class Codes: C07K 14/705 : Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/70578 : NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 14/7151 : for tumor necrosis factor [TNF], for lymphotoxin [LT]
• CPC Class Codes: C07K 16/18 : against material from animals or humans
• CPC Class Codes: C07K 16/22 : against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/241 : Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 : Interleukins [IL]
• CPC Class Codes: C07K 16/2803 : against the immunoglobulin superfamily
• CPC Class Codes: C07K 16/2809 : against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2818 : against CD28 or CD152
• CPC Class Codes: C07K 16/2863 : against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2875 : against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 16/2887 : against CD20
• CPC Class Codes: C07K 16/32 : against translation products of oncogenes
• CPC Class Codes: C07K 16/468 : Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/21 : from primates, e.g. man
• CPC Class Codes: C07K 2317/24 : containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/31 : multispecific
• CPC Class Codes: C07K 2317/524 : CH2 domain
• CPC Class Codes: C07K 2317/526 : CH3 domain
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/622 : Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/64 : comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/71 : Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/732 : Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 2319/32 : fusions with soluble part of a cell surface receptor, "decoy receptors"
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies

Abstract

The present disclosure provides various molecular constructs having a targeting element and/or an effector element. Methods for treating various diseases using such molecular constructs are also disclosed.

Claims

1. A linker unit comprising, a center core, a plurality of linking arms, and optionally a coupling arm, wherein,

the center core comprises, (1) a first polypeptide comprising a plurality of lysine (K) residues, wherein each K residue and its next K residue are separated by a filler sequence comprising glycine (G) and serine (S) residues, and the number of K residues ranges from 2 to 15;

or (2) a second polypeptide comprising the sequence of (X_aa-K)_n, where X_aa is a PEGylated amino acid having 2 to 12 repeats of ethylene glycol (EG) unit, and n is an integral from 2 to 15;

the plurality of linking arms are respectively linked to the K residues of the center core,

each of the plurality of linking arms has a maleimide group at its free terminus; and

the amino acid residue at the N- or C-terminus of the center core has an azide or an alkyne group;

or the amino acid residue at the N- or C-terminus of the center core is a cysteine residue, and the thiol group of the cysteine residue is linked with the coupling arm having the azide, the alkyne, a tetrazine or a strained alkyne group at the free terminus of the coupling arm.

2. The linker unit of claim 1, wherein the filler sequence has the sequence of GS, GGS, GSG, or SEQ ID NOs:

1-16.

3. The linker unit of claim 1, wherein the first polypeptide comprises 2-15 units of the sequence of G_1-5SK.

4. The linker unit of claim 3, wherein the first polypeptide comprises the sequence of (GSK)_2-15.

5. The linker unit of claim 1, wherein each of the linking arms is a PEG chain having 2-20 repeats of EG units.

6. The linker unit of claim 1, wherein the coupling arm is a PEG chain having 2-12 repeats of EG units.

7. The linker unit of claim 1, wherein the amino acid residue having the azide group is L-azidohomoalanine (AHA), 4-azido-L-phenylalanine, 4-azido-D-phenylalanine, 3-azido-L-alanine, 3-azido-D-alanine, 4-azido-L-homoalanine, 4-azido-D-homoalanine, 5-azido-L-ornithine, 5-azido-d-ornithine, 6-azido-L-lysine, or 6-azido-D-lysine.

8. The linker unit of claim 1, wherein the amino acid residue having the alkyne group is L-homopropargylglycine (L-HPG), D-homopropargylglycine (D-HPG), or beta-homopropargylglycine (β

-HPG).

9. The linker unit of claim 1, wherein the strained alkyne group is trans-cyclooctene (TCO), dibenzocyclooctyne (DBCO), difluorinated cyclooctyne (DIFO), bicyclononyne (BCN), or dibenzocyclooctyne (DICO).

10. The linker unit of claim 1, wherein the tetrazine group is 1,2,3,4-tetrazine, 1,2,3,5-tetrazine or 1,2,4,5-tetrazine, or derivatives thereof.

11. The linker unit of claim 1, further comprising a plurality of first elements that are respectively linked to the plurality of linking arms via thiol-maleimide reaction.

12. The linker unit of claim 11, further comprising a second element that is,

linked to the azide group via copper catalyzed azide-alkyne cycloaddition (CuAAC) reaction or strained-promoted azide-alkyne click chemistry (SPAAC) reaction;

linked to the alkyne group via CuAAC reaction;

linked to the strained alkyne group via inverse electron demand Diels-Alder (iEDDA) reaction or SPAAC reaction;

or

linked to the tetrazine group via iEDDA reaction.

13. The linker unit of claim 12, wherein the second element is linked to the azide or alkyne group at the N- or C-terminus of the center core via CuAAC reaction.

14. The linker unit of claim 13, further comprising a third element that is linked to the coupling arm via iEDDA reaction.

15. The linker unit of claim 14, wherein the third element is a long PEG chain having a molecular weight of about 20,000 to 50,000 daltons.

16. The linker unit of claim 12, wherein the second element is linked to the azide group at the N- or C-terminus of the center core via SPAAC reaction.

17. The linker unit of claim 16, further comprising a third element that is linked to the coupling arm via iEDDA reaction.

18. The linker unit of claim 17, wherein the third element is a long PEG chain having a molecular weight of about 20,000 to 50,000 daltons.

19. The linker unit of claim 12, wherein,

the first element is a first single-chain variable fragment (scFv) specific for a cytokine or a receptor of the cytokine;

or a soluble receptor of the cytokine; and

the second element is a second scFv specific for a tissue-associated extracellular matrix protein.

20. The linker unit of claim 19, wherein the tissue-associated extracellular matrix protein is selected from the group consisting of α

-aggrecan, collagen I, collagen II, collagen III, collagen V, collagen VII, collagen IX, and collagen XI.

21. The linker unit of claim 19, wherein the cytokine is selected from the group consisting of tumor necrosis factor-α

(TNF-α

), interleukin-17 (IL-17), IL-1, IL-6, shared protein of IL-12 and IL-23, and B cell activating factor (BAFF).

22. The linker unit of claim 19, wherein the receptor of the cytokine is a receptor specific for IL-6 (IL-6R) or a receptor specific for IL-17 (IL-17R).

23. The linker unit of claim 19, wherein the soluble receptor of the cytokine is specific for TNF-α

or IL-1.

24. The linker unit of claim 12, wherein,

the first element is a first scFv specific for a first cell surface antigen;

and

the second element is a second scFv specific for a second cell surface antigen.

25. The linker unit of claim 24, wherein the first cell surface antigen is selected from the group consisting of, CD5, CD19, CD20, CD22, CD23, CD27, CD30, CD33, CD34, CD37, CD38, CD43, CD72a, CD78, CD79a, CD79b, CD86, CD134, CD137, CD138, and CD319.

26. The linker unit of claim 24, wherein the second cell surface antigen is CD3 or CD16a.

27. The linker unit of claim 12, wherein,

the first element is a peptide hormone, a growth factor, or a first scFv specific for a tumor-associated antigen;

and

the second element is a second scFv specific for a cell surface antigen.

28. The linker unit of claim 27, wherein the peptide hormone is secretin, cholecystokinin (CCK), somatostatin, or thyroid-stimulating hormone (TSH).

29. The linker unit of claim 27, wherein the growth factor is selected from the group consisting of epidermal growth factor (EGF), mutant EGF, epiregulin, heparin-binding epidermal growth factor (HB-EGF), vascular endothelial growth factor A (VEGF-A), basic fibroblast growth factor (bFGF), and hepatocyte growth factor (HGF).

30. The linker unit of claim 27, wherein the tumor-associated antigen is selected from the group consisting of human epidermal growth factor receptor (HER1), HER2, HER3, HER4, carbohydrate antigen 19-9 (CA 19-9), carbohydrate antigen 125 (CA 125), carcinoembryonic antigen (CEA), mucin 1 (MUC 1), ganglioside GD2, melanoma-associated antigen (MAGE), prostate-specific membrane antigen (PSMA), prostate stem cell antigen (PSCA), mesothelin, mucine-related Tn, Sialyl Tn, Globo H, stage-specific embryonic antigen-4 (SSEA-4), and epithelial cell adhesion molecule (EpCAM).

31. The linker unit of claim 27, wherein the cell surface antigen is CD3 or CD16a.

32. The linker unit of claim 12, wherein,

the first element is a first scFv specific for ligand of receptor activator of nuclear factor κ

B (RANKL); and

the second element is a second scFv specific for collagen I or osteonectin.

33. The linker unit of claim 12, wherein,

the first element is an scFv specific for VEGF-A;

and

the second element is a long PEG chain having a molecular weight of about 20,000 to 50,000 daltons.