Molecular constructs with targeting and effector moieties for treating central nervous system diseases

• US 20160339115A1
• Filed: 05/20/2016
• Published: 11/24/2016
• Est. Priority Date: 05/20/2015
• Status: Active Grant

• Current Assignee: Immunwork Inc.
• Original Assignee: Immunwork Inc.
• Inventors: CHANG, Tse-Wen, CHU, Hsing-Mao
• Granted Patent: US 10,202,455 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• 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 arm linked to the first center core,

optionally, a first connecting arm linked to the first linking arm,

a first element linked to the first linking arm or the first connecting arm, and

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

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,

optionally, a second connecting arm linked to the second linking arm,

a second element linked to the second linking arm or the second connecting arm, and

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

wherein,

the first and second linker units are coupled to each other via CuAAC reaction, SPAAC reaction or 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 element is an scFv specific for transferrin receptor, and the second element is interferon-β

(IFN-β

), fingolimod, fingolimod phosphate, or an scFv specific for integrin α

4 or β

-amyloid.

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 2, further comprising a plurality of first and second connecting arms respectively linked to the first and second linking arms.

5. The molecular construct of claim 4, further comprising a plurality of first and second elements respectively linked to the first and second connecting arms.

6. The molecular construct of claim 1, wherein,

each of the first and second linking arms is a PEG chain having 2-20 repeats of EG units or a PEG chain having 2-20 repeats of EG units with a disulfide linkage at the free terminus thereof;

and

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

7. The molecular construct of claim 4, wherein each of the first and second connecting arms is the PEG chain having 2-20 repeats of EG units or the PEG chain having 2-20 repeats of EG units with a disulfide linkage at the terminus that is not linked with the linking arm.

8. The molecular construct of claim 1, wherein,

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;

and

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

9. The molecular construct of claim 8, wherein the strained alkyne group is DBCO, DIFO, BCN, or DICO.

10. The molecular construct of claim 1, wherein,

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;

and

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

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

12. The molecular construct of claim 1, wherein at least one of the first and the second center cores is a compound core, wherein the 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 strained alkyne or a tetrazine group at the free-terminus thereof.

13. The molecular construct of claim 12, wherein the compound 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.

14. The molecular construct of claim 12, wherein,

one of the first and the second center cores is the compound core, wherein the coupling arm linked to the compound core is linked via forming an amide bond with one of the plurality of amine groups of the compound core and 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 a polypeptide 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.

15. The molecular construct of claim 1, wherein at least one of the first and second center cores is a polypeptide comprising a plurality of lysine (K) residues.

16. The molecular construct of claim 15, wherein the number of K residues ranges from 2 to 15, and each K residue and its next K residue are separated by a filler sequence comprising glycine (G) and serine (S) residues.

17. The molecular construct of claim 16, wherein the filler sequence has the sequence of GS, GGS, GSG, or SEQ ID NOs:

1-16.

18. The molecular construct of claim 16, wherein the polypeptide comprises 2-15 units of the sequence of G_1-5SK.

19. The molecular construct of claim 18, wherein the polypeptide comprises the sequence of (GSK)_2-15.

20. The molecular construct of claim 15 wherein the polypeptide comprises 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.

21. The molecular construct of claim 20, wherein both the first and second center cores are the polypeptides.

22. The molecular construct of claim 21, wherein the N-terminus of each of the first and second center cores is modified with an acetyl group.

23. The molecular construct of claim 21, 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-HPG, D-HPG, or β

-HPG; and

the first and second center cores are coupled to each other via CuAAC reaction occurred between the N- or C-terminal amino acid residues.

24. The molecular construct of claim 21, wherein each of the first and second center cores comprises a cysteine residue at the N- or C-terminus thereof;

and the first and second coupling arms are respectively linked to the cysteine residues of the first and second center cores via thio-maleimide reaction.

25. The molecular construct of claim 21, 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 that is linked to the cysteine residue is linked via thio-maleimide reaction and 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.

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

27. The molecular construct of claim 26, 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.

28. The molecular construct of claim 27, wherein the third element is different from the first element and the second element.

29. A method of treating a subject having or suspected of having a CNS disease, comprising administering to the subject in need thereof a therapeutically effective amount of the molecular construct of claim 1.

30. The method of claim 29, wherein,

the CNS disease is multiple sclerosis,

the first element is the scFv specific for transferrin receptor, and

the second element is IFN-β

, fingolimod, fingolimod phosphate, or the scFv specific for integrin α

4.

31. The method of claim 29, wherein,

the CNS disease is Alzheimer'"'"'s disease,

the first element is the scFv specific for transferrin receptor, and

the second element is the scFv specific for β

-amyloid.