Fabs-in-tandem immunoglobulin and uses thereof

• US 10,266,608 B2
• Filed: 05/04/2016
• Issued: 04/23/2019
• Est. Priority Date: 12/30/2013
• Status: Active Grant

• Current Assignee: EpimAb Biotherapeutics Inc. (Shanghai EpimAb Biotherapeutics Co. Ltd.)
• Original Assignee: EpimAb Biotherapeutics Inc. (Shanghai EpimAb Biotherapeutics Co. Ltd.)
• Inventors: Wu, Chengbin
• Primary Examiner(s): Wu, Julie
• Application Number:
  US15/146,540
• Publication Number:
  US 20160289343A1
• Time in Patent Office: 1,084 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• 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 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/02 Nasal agents, e.g. decongestants
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• 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 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• 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 29/00 : Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 3/00 : Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 33/00 : Antiparasitic agents
• 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: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 5/00 : Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• 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 16/2887 : against CD20
• 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 2317/31 : multispecific
• CPC Class Codes: C07K 2317/35 : Valency
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/522 : CH1 domain
• CPC Class Codes: C07K 2317/55 : Fab or Fab'
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/64 : comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/66 : comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
• CPC Class Codes: C07K 2317/76 : Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/90 : characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
• CPC Class Codes: C07K 2317/92 : Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide

Abstract

The present invention provides multivalent and multispecific binding proteins that are capable of binding two or more antigens, or two or more epitopes. The present invention also provides methods of making and using such multivalent and multispecific binding proteins, including methods of using such binding proteins for prevention or treatment of various diseases, or for detecting specific antigens in vitro or in vivo.

Claims

1. A composition comprising a binding protein, said binding protein comprising three polypeptide chains,

wherein the first polypeptide chain comprises, from amino terminus to carboxyl terminus, either (i) VL_A-CL-VH_B-CH1-Fc, wherein CL is fused directly to VH_B, or (ii) VH_B-CH1-VL_A-CL-Fc, wherein CH1 is fused directly to VL_A,

wherein the second polypeptide chain comprises, from amino terminus to carboxyl terminus, VH_A-CH1,

wherein the third polypeptide chain comprises, from amino terminus to carboxyl terminus, VL_B-CL,

wherein A is a first epitope or antigen, and B is a second epitope or antigen, and wherein A and B are different epitopes of the same antigen or are different antigens;

wherein VL_A is a light chain variable domain of a first parental antibody that binds A, CL is an antibody light chain constant domain, VH_B is a heavy chain variable domain of a second parental antibody that binds B, CH1 is a first constant domain of an antibody heavy chain, VH_A is a heavy chain variable domain of said first parental antibody that binds A, and VL_B is a light chain variable domain of said second parental antibody that binds B;

wherein said binding protein binds to both A and B;

wherein the binding protein comprises two of said first polypeptide chains, two of said second polypeptide chains, and two of said third polypeptide chains under non-reducing conditions; and

wherein 90% or more of the binding protein in the composition is a single, monomeric, tetravalent and bispecific antibody as determined by size exclusion chromatography.

2. The composition of claim 1, wherein the binding protein comprises:

3. The composition of claim 1, wherein the Fc region of said binding protein is the Fc of human IgG1.

4. The composition of claim 1, wherein the Fc region of said binding protein has the amino acid sequence of SEQ ID NO:

20.

5. The composition of claim 1, wherein the binding protein is capable of binding pairs of targets selected from the group consisting of IL-1α

and IL-1β

;

IL-12 and IL-18, TNFα and

IL-23, TNFα and

IL-13;

TNF and IL-18;

TNF and IL-12;

TNF and IL-1beta;

TNF and MIF;

TNF and IL-6, TNF and IL-6 Receptor, TNF and IL-17;

IL-17 and IL-20;

IL-17 and IL-23;

TNF and IL-15;

TNF and VEGF;

VEGFR and EGFR;

IL-13 and IL-9;

IL-13 and IL-4;

IL-13 and IL-5;

IL-13 and IL-25;

IL-13 and TARC;

IL-13 and MDC;

IL-13 and MIF;

IL-13 and TGF-β

;

IL-13 and LHR agonist;

IL-13 and CL25;

IL-13 and SPRR2a;

IL-13 and SPRR2b;

IL-13 and ADAMS; and

TNFα and

PGE4, IL-13 and PED2, TNF and PEG2, CD138 and CD20;

CD138 and CD40;

CD19 and CD20;

CD19 and CD22;

CD20 and CD3;

CD3 and CD33;

CD16 and CD33, CD3 and CD133;

CD38 &

CD138;

CD38 and CD20;

CD20 and CD22;

CD38 and CD40;

CD40 and CD20;

CD-8 and IL-6;

CSPGs and RGM A;

IGF1 and IGF2;

IGF1/2 and Erb2B;

IGF-1R and EGFR;

EGFR and CD13;

IGF-1R and ErbB3;

EGFR-2 and IGFR;

VEGFR-2 and c-Met;

VEGF-A and Angiopoietin-2 (Ang-2);

IL-12 and TWEAK;

IL-13 and IL-1beta;

MAG and RGM A;

NgR and RGM A;

NogoA and RGM A;

OMGp and RGM A;

CTLA-4 and BTNO2;

PD-1 and CTLA-4;

PD-L1 and CTLA-4;

LAG-3 and CTLA-4;

ICOS and CTLA-4;

PD-1 and TIM-3;

CD137 and CD20;

CD137 and EGFR;

CD137 and Her-2;

CD137 and PD-1;

CD137 and PD-L1;

CD137 and CD27;

CD137 and GITR;

CD137 and HVEM;

CD137 and OX40;

CD27 and HVEM;

CD137 and LAG-3;

CD137 and TIM-3;

CD137 and VISTA;

c-Met and PD-L1;

Lag-3 and PD-L1;

TIM-3 and PD-L1;

EGFR and PD-L1;

ICOS and PD-L1;

Her2 and PD-L1;

PD-L1 and PD-L2;

EGFR and PD-1;

PD-1 and PD-L1;

OX40 and PD-L1;

VEGF and PD-L1;

LAG-3 and TIM-3;

OX40 and PD-1;

BTLA and PD-1;

VISTA and PD-1;

TIM3 and PD-1;

LAG-3 and PD-1;

LAG-3 and Her-2;

LAG-3 and EGFR;

LAG-3 and BTLA;

LAG-3 and VISTA;

OX40 and GITR;

OX40 and CD27;

OX40 and HVEM;

GITR and HYEM;

EGFR and DLL-4;

VEGF and DLL-4;

PDGFR and VEGF;

EpCAM and CD3;

Her2 and CD3;

CD19 and CD3;

CD19 and CD20;

CEA and EGFR;

EpCAM and EGFR;

PSMA and PSCA;

EphA2 and Her2, EpCAM and CD3;

EGFR and Her3;

CD16a and CD30;

CD30 and PSMA;

EGFR and CD3;

CEA and CD3;

TROP-2 and HSG;

TROP-2 and CD3;

VEGF and EGFR;

HGF and VEGF;

VEGF and VEGF (same or a different epitope);

VEGF and HGF;

EGFR and cMet;

PDGF and VEGF;

ICOS and PD-1, Lag-3 and PD-1, CD47 and CD20, RGM A and RGM B;

Te38 and TNFα

;

TNFα and

Blys;

TNFα and

CD-22;

TNFα and

CTLA-4 domain;

TNFα and

GP130;

TNFα and

IL-12p40; and

TNFα and

RANK ligand.

6. The composition of claim 1, wherein the binding protein is capable of binding one or more epitopes on CTLA-4 and/or one or more epitopes on PD-1.

7. A pharmaceutical composition comprising the composition according to claim 1 and one or more pharmaceutically acceptable carriers.

8. The pharmaceutical composition of claim 7, wherein the composition further comprises at least one additional therapeutic agent selected from the group consisting of a cytotoxic agent, an angiogenesis inhibitor, a kinase inhibitor, a co-stimulation molecule blocker;

an adhesion molecule blocker, an anti-cytokine antibody or functional fragment thereof, methotrexate, cyclosporin, rapamycin, FK506, a detectable label or reporter, a TNF antagonist, an antirheumatic, a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteroid, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.

9. The composition of claim 1, wherein the binding protein is conjugated to an agent selected from the group consisting of an immunoadhesion molecule, an imaging agent, a therapeutic agent, and a cytotoxic agent.

10. The composition according to claim 9, wherein the imaging agent is selected from the group consisting of a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, and biotin.

11. A diagnostic kit comprising the composition according to claim 1.

12. The composition according to claim 5, wherein the binding protein is capable of binding cMet and EGFR.