Pharmaceuticals in joint-linker configurations for treating pathological blood clots

• US 10,383,947 B2
• Filed: 07/18/2016
• Issued: 08/20/2019
• 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): Halvorson, Mark
• Assistant Examiner(s): Akhoon, Kauser M
• Application Number:
  US15/212,301
• Publication Number:
  US 20170056518A1
• Time in Patent Office: 1,128 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 47/68033 the drug being a maytansine
• 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 an effector element. Methods for treating various diseases using such molecular constructs are also disclosed.

Claims

1. A molecular construct comprising a first linker unit and a second linker unit, wherein,

the first linker unit comprises,

a first center core comprising a plurality of amine groups,

a first linking arm linked to the first center core,

a first element linked to the first linking arm, and

optionally, a first coupling arm linked to the first center core;

the second linker unit comprises,

a second center core comprising a plurality of amine groups,

a second linking arm linked to the second center core,

a second element linked to the second linking arm, and

optionally, a second coupling arm linked to the second center core; and

the first and second linker units are coupled to each other via copper catalyzed azide-alkyne cycloaddition (CuAAC) reaction, strained-promoted azide-alkyne click chemistry (SPAAC) reaction or inverse electron demand Diels-Alder (iEDDA) reaction occurred between any of the followings;

the first and second center cores, the first coupling arm and the second center core, the first and second coupling arms, or the first center core and the second coupling arm;

wherein,

the first or second center core is independently a polypeptide core or a compound core, wherein

the polypeptide core comprises, (1) 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) the sequence of (Xaa-K)n, where Xaa is a PEGylated amino acid having 2 to 12 repeats of ethylene glycol (EG) unit, and n is an integer from 2 to 15;

wherein at least one of the N- and C-terminal amino acid residues of the polypeptide is an amino acid having an azide or an alkyne group or is a cysteine residue, and when one of the N- and C-terminal amino acid residues is the cysteine residue, the linker unit further comprises, optionally, the coupling arm that is linked to the cysteine residue via the thiol group of the cysteine residue and has an azide, an alkyne, a tetrazine, a cyclooctene, or a cyclooctyne group at the free terminus thereof; and

the compound core is selected from the group consisting of, benzene-1,3,5-triamine, 2-(aminomethyl)-2-methylpropane-1,3-diamine, tris(2-aminoethyl)amine, benzene-1,2,4,5-tetraamine, 3,3′

,5,5′

-tetraamine-1,1′

-biphenyl, tetrakis(2-aminoethyl)methane, tetrakis-(ethylamine)hydrazine, N,N,N′

,N′

-tetrakis(aminoethyl)ethylenediamine, benzene-1,2,3,4,5,6-hexaamine, 1-N,1-N,3-N,3-N,5-N,5-N-hexakis(methylamine)-benzene-1,3,5-triamine, 1-N,1-N,2-N,2-N,4-N,4-N,5-N,5-N,-octakis(methylamine)-benzene-1,2,4,5-triamine, and N,N-bis[(1-amino-3,3-diaminoethyl)pentyl] methanediamine; and

the first or second coupling arm linked to said compound core is linked thereto via forming an amide bond with one of the plurality of amine groups of the compound core and has an azide, an alkyne, a tetrazine, a cyclooctene, or a cyclooctyne group at the free-terminus thereof;

the first and second linking arms are respectively linked to the amine groups of the first and second center cores via forming an amide bond;

the first and second elements are respectively linked to the first and second linking arms via forming an amide bond, or via thiol-maleimide, CuAAC, iEDDA, or SPAAC reaction; and

the first element is an scFv specific for fibrin, and the second element is a tissue plasminogen activator, an inhibitor of Factor Xa, or an inhibitor of thrombin.

2. The molecular construct of claim 1, wherein the first and second linker units respectively comprise a plurality of the first and second linking arms linked thereto.

3. The molecular construct of claim 2, further comprising a plurality of first and second elements respectively linked to the first and second linking arms.

4. The molecular construct of claim 1, wherein,

each of the first and second linking arms is a PEG chain having 2-20 repeats of ethylene glycol (EG) units;

and

each of the first and second coupling arms is a PEG chain having 2-12 repeats of EG units.

5. The molecular construct of claim 1, wherein,

one of the first and second coupling arms has the azide group at the free-terminus thereof, and the other of the first and second coupling arms has the alkyne or the cyclooctyne group at the free-terminus thereof;

and

the first and second linker units are coupled to each other via copper catalyzed azide-alkyne cycloaddition (CuAAC) reaction or strained-promoted azide-alkyne click chemistry (SPAAC) reaction occurred between the first and second coupling arms.

6. The molecular construct of claim 1, wherein the cyclooctyne group is dibenzocyclooctyne (DBCO), difluorinated cyclooctyne (DIFO), bicyclononyne (BCN), or dibenzocyclooctyne (DICO).

7. The molecular construct of claim 1, wherein,

one of the first and second coupling arms has the tetrazine group at the free-terminus thereof, and the other of the first and second coupling arms has the cyclooctene group at the free-terminus thereof;

and

the first and second linker units are coupled to each other via iEDDA reaction occurred between the first and second coupling arms.

8. The molecular construct 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.

9. The molecular construct of claim 1, wherein one of the first and the second center cores is the compound core.

10. The molecular construct of claim 1, wherein,

one of the first and the second center cores is the compound core, and the coupling arm linked to the compound core has a DBCO, a DIFO, a BCN, or a DICO group at the free-terminus thereof;

the other of the first and the second center cores is the polypeptide core in which the N- or C-terminal amino acid residue 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; and

the first and second linker units are coupled to each other via SPAAC reaction occurred between the coupling arm and the N- or C-terminal amino acid residue.

11. The molecular construct of claim 1, wherein at least one of the first and second center cores is the polypeptide core.

12. The molecular construct of claim 1, wherein the filler sequence has the sequence of GS, GGS, GSG, or SEQ ID NOs:

1-16.

13. The molecular construct of claim 1, wherein the polypeptide core comprises 2-15 units of the sequence of G_1-5SK.

14. The molecular construct of claim 13, wherein the polypeptide core comprises the sequence of (GSK)_2-15.

15. The molecular construct of claim 1, wherein the first and second center cores are independently the polypeptide cores.

16. The molecular construct of claim 15, wherein the N-terminus of each of the first and second center cores is modified with an acetyl group.

17. The molecular construct of claim 15, wherein,

the N- or C-terminal amino acid residue of one of the first and second center cores is 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;

the N- or C-terminal amino acid residue of the other of the first and second center cores is L-homopropargylglycine (L-HPG), D-homopropargylglycine (D-HPG), or beta-homopropargylglycine (β

-HPG); and

the first and second center cores are coupled to each other via copper catalyzed azide-alkyne cycloaddition (CuAAC) reaction occurred between the N- or C-terminal amino acid residues.

18. The molecular construct of claim 15, wherein each of the first and second center cores comprises a cysteine residue at the N- or C-terminus thereof.

19. The molecular construct of claim 15, wherein,

the N- or C-terminal amino acid residue of one of the first and second center cores is 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;

the N- or C-terminal amino acid residue of the other of the first and second center cores is a cysteine residue, and the coupling arm has a DBCO, a DIFO, a BCN, or a DICO group at the free-terminus thereof; and

the first and second center cores are coupled to each other via SPAAC reaction occurred between the N- or C-terminal amino acid residue and the coupling arm.

20. The molecular construct of claim 1, wherein the tissue plasminogen activator is alteplase, reteplase, tenecteplase, or lanoteplase.

21. The molecular construct of claim 1, wherein the inhibitor of Factor Xa is apixaban, edoxaban, or rivaroxaban;

and the inhibitor of thrombin is argatroban or melagatran.

22. The molecular construct of claim 1, further comprising a third linking arm linked to the first or the second linker unit.

23. The molecular construct of claim 22, wherein the third linking arm has a maleimide group at the free terminus thereof, and a third element is linked to the third linking arm via thiol-maleimide reaction.

24. The molecular construct of claim 23, wherein the third element is different from the first element and the second element.

25. The molecular construct of claim 24, wherein the third element is a long PEG chain having a molecular weight of about 20,000 to 50,000 Daltons.