ANTIBODY LOCKER FOR THE INACTIVATION OF PROTEIN DRUG

• US 20160185875A1
• Filed: 05/28/2014
• Published: 06/30/2016
• Est. Priority Date: 05/28/2013
• Status: Active Grant

• Current Assignee: DCB-USA LLC, Kaohsiung Medical University
• Original Assignee: DCB-USA LLC
• Inventors: Chuang, Chih-Hung, Ko, Hsiu-Fen, Lu, Yun-Chi, Cheng, Tian-Lu
• Granted Patent: US 10,633,453 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 16/241 Tumor Necrosis Factors
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 16/245 IL-1
• 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 16/2866 against receptors for cytokines, lymphokines, interferons
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• 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 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Abstract

Disclosed herein is a hinge antibody capable of being selectively activated in a target cell or tissue to treat a condition therein. The hinge antibody includes a functional antibody, two inhibitory domains and four cleavable linkers. The functional antibody is capable of treating the condition in an activated state, and has two light chains and two heavy chains. Each inhibitory domain includes a hinge domain of an immunoglobulin and consists of two peptide arms. Each cleavable linker includes a peptide substrate cleavable by an enzyme specifically or highly expressed in the target cell or tissue, and connects one of the peptide arms of the inhibitory domains to the N-terminal of one of the light chains and heavy chains of the functional antibody. Also disclosed herein are methods for preparing and using this hinge antibody.

Claims

1. A hinge antibody capable of being selectively activated in a target cell or tissue to treat a condition therein, comprising:

a functional antibody capable of treating the condition in an activated state, comprising two light chains and two heavy chains;

two inhibitory domains, wherein each inhibitory domain consists of two peptide arms interconnected by disulfide bonds; and

four cleavable linkers, wherein each cleavable linker comprises a peptide substrate cleavable by an enzyme that is specifically or highly expressed in the target cell or tissue, and connects one of the two peptide arms of the two inhibitory domains to the N-terminals of one of the two light chains and two heavy chains of the functional antibody.

2. The hinge antibody of claim 1, wherein the functional antibody is selected from the group consisting of, anti-TNF-α

antibody, anti-RANKL antibody, anti-CTLA-4 antibody, anti-HER2 antibody, anti-EGFR antibody, anti-VEGF antibody, anti-VEGFR2) antibody, anti-IL6R antibody, anti-IL12/23 antibody, anti-CD3 antibody, anti-CD11a antibody, anti-CD20 antibody, anti-CD25 antibody, anti-CD30 antibody, anti-CD33 antibody and anti-CD52 antibody

3. The hinge antibody of claim 1, wherein the light chain of the functional antibody has any of the amino acid sequences of SEQ ID Nos. 1, 2, 3, 4, 5, 6, 7, 8 or 9, and the heavy chain of the functional antibody has any of the amino acid sequences of SEQ ID Nos. 58, 59, 60, 61, 62, 63, 64, 65 and 66.

4. The hinge antibody of claim 1, wherein each of the two inhibitory domains is a hinge domain of an immunoglobulin A (IgA), immunoglobulin D (IgD) or an immunoglobulin G (IgG), or a fragment of the hinge domain.

5. The hinge antibody of claim 4, wherein the IgA is IgA1 or IgA2.

6. The hinge antibody of claim 4, wherein the IgG is IgG1, IgG2, IgG3 or IgG4.

7. The hinge antibody of claim 4, wherein each of the inhibitory domains comprises any of the amino acid sequences of SEQ ID Nos. 10, 11, 12, 13, 14, 15, 54 or 55.

8. The hinge antibody of claim 1, wherein the peptide substrate is cleavable by any of the following enzyme:

a matrix metalloproteinase (MMP), a cathepsin (CTS), a caspase (CASP), or a disintegrin and metalloproteinase (ADAM).

9. The hinge antibody of claim 8, wherein the enzyme is MMP-2 or MMP-9 and each cleavable linker comprises the amino acid sequence of SEQ ID No. 16.

10. The hinge antibody of claim 1, wherein,

the functional antibody is an anti-TNF-α

antibody, wherein the light chain thereof comprises the amino acid sequence of SEQ ID No. 1, and the heavy chain thereof comprises the amino acid sequence of SEQ ID No. 58;

each of the cleavable linkers comprises the amino acid sequence of SEQ ID No. 16; and

each of the inhibitory domains comprises the amino acid sequence of SEQ ID No. 10.

23. A method for treating cancer or an autoimmune disease in a subject, comprising administering to the subject a therapeutically effective amount of the hinge antibody of any of claims 1 to 10.

24. The method of claim 23, wherein the hinge antibody is administered subcutaneously, orally, intravenously, intrathecally or intramuscularly.

11. An expression system for producing a hinge antibody capable of being selectively activated in a target cell or tissue to treat a condition therein, comprising,

a first nucleic acid sequence, comprising, from 5′

to 3′

,

a first inhibitory domain-encoding region encoding a first peptide arm of an inhibitory domain of the hinge antibody, wherein the inhibitory domain comprises a hinge domain of an immunoglobulin, or a fragment of the hinge domain,

a first cleavable linker-encoding region encoding a cleavable linker of the hinge antibody, wherein the cleavable linker is a peptide substrate cleavable by an enzyme that is specifically or highly expressed in the target cell or tissue, and

a light chain-encoding region encoding a light chain of a functional antibody of the hinge antibody, wherein the functional antibody is capable of treating the condition in an activated state; and

a second nucleic acid sequence, comprising, from 5′

to 3′

,

a second inhibitory domain-encoding region encoding a second peptide arm of the inhibitory domain of the hinge antibody,

a second cleavable linker-encoding region encoding the cleavable linker of the hinge antibody, and

a heavy chain-encoding region encoding a heavy chain of the functional antibody of the hinge antibody.

12. The expression system of claim 11, further comprising a connecting nucleic acid sequence connecting the first nucleic acid sequence and the second nucleic acid sequence, wherein the connecting nucleic acid sequence is a Furin-2A-encoding sequence or an internal ribosome entry site (IRES) sequence.

13. The expression system of claim 12, further comprising a regulatory sequence operably linked to the first nucleic acid sequence and the second nucleic acid sequence to express the first nucleic acid sequence, the second nucleic acid sequence and the connecting nucleic acid sequence.

14. The expression system of claim 11, further comprising,

a first regulatory sequence operably linked to the first nucleic acid sequence to express the first nucleic acid sequence;

and

a second regulatory sequence operably linked to the second nucleic acid sequence to express the second nucleic acid sequence.

15. The expression system of claim 14, wherein,

the first nucleic acid sequence and first regulatory sequence are constructed in a first expression vector;

and

the second nucleic acid sequence and second regulatory sequence are constructed in a second expression vector.

16. The expression system of claim 14, wherein the first nucleic acid sequence, the first regulatory sequence, the second nucleic acid sequence, and the second regulatory sequence are constructed in a single expression vector.

17. The expression system of claim 11, wherein the functional antibody is selected from the group consisting of, anti-TNF-α

antibody, anti-RANKL antibody, anti-CTLA-4 antibody, anti-HER2 antibody, anti-EGFR antibody, anti-VEGF antibody, anti-VEGFR2) antibody, anti-IL6R antibody, anti-IL12/23 antibody, anti-CD3 antibody, anti-CD11a antibody, anti-CD20 antibody, anti-CD25 antibody, anti-CD30 antibody, anti-CD33 antibody and anti-CD52 antibody.

18. The expression system of claim 11, wherein the immunoglobulin is an immunoglobulin A (IgA), an immunoglobulin D (IgD), or an immunoglobulin G (IgG).

19. The synthetic nucleic acid molecule of claim 18, wherein the IgA is IgA1 or IgA2.

20. The synthetic nucleic acid molecule of claim 18, wherein the IgG is IgG1, IgG2, IgG3 or IgG4.

21. The synthetic nucleic acid molecule of claim 11, wherein the peptide substrate is cleavable by any of the following enzyme:

a matrix metalloproteinase (MMP), a cathepsin (CTS), a caspase (CASP), or a disintegrin and metalloproteinase (ADAM).

22. The synthetic nucleic acid molecule of claim 11, wherein the synthetic nucleic acid molecule has a nucleotide sequence that is any of SEQ ID Nos. 17 to 50.