Molecular constructs with targeting and effector moieties comprising an SCFV specific for ectodomain of transferrin-1 receptor and fingolimod

• US 10,202,455 B2
• Filed: 05/20/2016
• Issued: 02/12/2019
• Est. Priority Date: 05/20/2015
• Status: Active Grant

• Current Assignee: Immunwork Inc.
• Original Assignee: Immunwork Inc.
• Inventors: Chang, Tse-Wen, Chu, Hsing-Mao
• Primary Examiner(s): Wang, Chang-Yu
• Application Number:
  US15/159,834
• Publication Number:
  US 20160339115A1
• Time in Patent Office: 998 Days
• CPC Class Codes: A61K 31/137 Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
• 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 47/64 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 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 47/6849 the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
• CPC Class Codes: A61K 47/6883 Polymer-drug antibody conjugates, e.g. mitomycin-dextran-Ab; DNA-polylysine-antibody complex or conjugate used for therapy
• 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 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C07K 14/565 IFN-beta
• CPC Class Codes: C07K 16/1027 Paramyxoviridae, e.g. respiratory syncytial virus
• CPC Class Codes: C07K 16/1203 from Gram-negative bacteria
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/283 against Fc-receptors, e.g. CD16, CD32, CD64 CD23 C07K16/2851
• CPC Class Codes: C07K 16/2839 against the integrin superfamily
• CPC Class Codes: C07K 16/2881 against CD71
• CPC Class Codes: C07K 17/02 Peptides being immobilised on, or in, an organic carrier
• CPC Class Codes: C07K 17/06 attached to the carrier via a bridging agent
• CPC Class Codes: C07K 2317/31 : multispecific
• CPC Class Codes: C07K 2317/35 : Valency
• 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 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2319/33 : fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 : containing domain for protein-protein interaction
• CPC Class Codes: C07K 2319/74 : containing a fusion for binding to a cell surface receptor
• CPC Class Codes: C07K 7/08 : having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68

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 atm linked to the first center core, and

a first element linked to the first linking arm;

and

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, and

a second element linked to the second linking arm,

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

wherein each of the first and second center core is a polypeptide that 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 in the polypeptide 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 integral from 2 to 15;

wherein one of the N- and C-terminal amino acid residues of the polypeptide is a cysteine residue;

wherein the first linker unit further comprises a first 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 a free terminus of the first coupling arm;

wherein the second linker unit further comprises a second coupling arm that is linked to the cysteine residue via the thiol group of the cysteine residue and has the azide, the alkyne, a tetrazine, a cyclooctene, or a cyclooctyne group at a free terminus of the second coupling arm;

wherein if one of the first and second coupling arms has an azide group at the free-terminus thereof, and the other of the first and second coupling arms has an alkyne or a cyclooctyne group at the free-terminus thereof, the first and second linker units are coupled to each other via CuAAC reaction or SPAAC reaction occurred between the first and second coupling amines;

wherein if one of the first and second coupling arms has a tetrazine group at the free-terminus thereof, and the other of the first and second coupling arms has a cyclooctene group at the free-terminus thereof, the first and second linker units are coupled to each other via iEDDA reaction occurred between the first and second coupling arms;

the first and second linking arms are respectively linked to another of the N- and C-terminal amino acid residues of the first and second center cores, wherein each of the first and second linking arms, at a terminus thereof, has maleimide, an azide, an alkyne, a tetrazine, a cyclooctene, or a cyclooctyne group or is a polyethylene glycol (PEG) chain having 2-20 repeats of ethylene glycol (EG) units or a PEG chain having 2-20 repeats of EG units with a disulfide linkage; and

the first element is an scFv specific for ectodomain of transferrin-1 receptor (TfR1), and the second element is fingolimod or fingolimod phosphate.

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

3. 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.

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

1-16.

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

6. The molecular construct of claim 5, wherein the polypeptide comprises the sequence of (GSK)_2-15.

7. The molecular construct of claim 1, wherein the first and second coupling arms are respectively linked to the cysteine residues of the first and second center cores via thio-maleimide reaction.

8. The molecular construct of claim 1, wherein the first element is linked to the first linking arm via forming an amide bond, or via thiol-maleimide, CuAAC, iEDDA, or SPAAC reaction.

9. The molecular construct of claim 1, wherein the second element is linked to the second linking arm via forming an amide bond, or via thiol-maleimide, CuAAC, iEDDA, or SPAAC reaction.