Anti-DLL4/VEGF dual variable domain immunoglobulin and uses thereof

• US 9,045,551 B2
• Filed: 06/11/2014
• Issued: 06/02/2015
• Est. Priority Date: 11/01/2012
• Status: Active Grant

• Current Assignee: Abbvie Incorporated
• Original Assignee: Abbvie Incorporated
• Inventors: Gu, Jijie, Ambrosi, Dominic J., Lappe, Susan E., Li, Yingchun, Hickson, Jonathan A., Haasch, Deanna L., Gupta, Supriya, Chari, Ravi, Zamiri, Camellia, Naumovski, Louie, Cao, Xianhua
• Primary Examiner(s): DAHLE, CHUN WU
• Application Number:
  US14/301,546
• Publication Number:
  US 20140348835A1
• Time in Patent Office: 356 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 31/337 having four-membered rings, e.g. taxol
• 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 31/495 having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazinesA61K31/48 takes precedence ; with three nitrogen atoms A61K31/53
• CPC Class Codes: A61K 31/513 having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
• CPC Class Codes: A61K 31/525 Isoalloxazines, e.g. riboflavins, vitamin B2
• CPC Class Codes: A61K 31/7068 having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
• CPC Class Codes: A61K 39/395 Antibodies agglutinins A61K38/36 ; as drug carriers A61K47/50; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/39591 Stabilisation, fragmentation
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/10 Antioedematous agents; Diuretics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 16/22 : against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/28 : against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: C07K 16/468 : Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• 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 2317/35 : Valency
• CPC Class Codes: C07K 2317/52 : Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 : variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/565 : Complementarity determining region [CDR]
• 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 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

Abstract

Disclosed herein are multivalent and multispecific binding proteins, methods of making the binding proteins, and their uses in the diagnosis, monitoring, inhibition, prevention and/or treatment of cancers, tumors, and/or other angiogenesis-dependent diseases characterized by aberrant DLL4 and/or VEGF expression or activity.

Claims

1. A binding protein comprising first and second polypeptide chains, each independently comprising VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first variable domain;

VD2 is a second variable domain;

C is a constant domain;

X1 is a linker;

X2 is an Fc region;

n is 0 or 1,

wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site, and wherein the binding protein is capable of binding DLL4 and VEGF, wherein the first polypeptide chain of the binding protein comprises SEQ ID NO;

56 and the second polypeptide chain of the binding protein comprises SEQ ID NO;

64.

2. The binding protein of claim 1, wherein the binding protein comprises a first polypeptide chain comprising a first VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first heavy chain variable domain;

VD2 is a second heavy chain variable domain;

C is a heavy chain constant domain;

X1 is a linker;

X2 is an Fc region;

n is 0 or 1, and

wherein the binding protein comprises a second polypeptide chain comprising a second VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first light chain variable domain;

VD2 is a second light chain variable domain;

C is a light chain constant domain;

X1 is a linker;

n is 0 or 1 for (X1)n;

n is 0 for (X2)n,

wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site.

3. The binding protein of claim 1, wherein the binding protein comprises two first and two second polypeptide chains and four functional target binding sites.

4. The binding protein of claim 1, wherein the binding protein is capable of binding:

(a) VEGF with a dissociation constant (K_D) of at most about 7.40×

10<sup>−

9</sup>M, as measured by surface plasmon resonance; and

/or

(b) DLL4 with a dissociation constant (K_D) of at most about 3.40×

10<sup>−

8</sup> M or 5.00×

10<sup>−

8</sup> M, as measured by surface plasmon resonance.

5. The binding protein of claim 1, wherein the Fc region of the binding protein is an Fc region from an IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD, or a variant thereof.

6. The binding protein of claim 1, wherein the Fc region of the binding protein is a variant sequence Fc region.

7. The binding protein of claim 1, wherein the binding protein comprises the constant region sequences from SEQ ID NO:

73 and/or SEQ ID NO;

74.

8. The binding protein of claim 1, wherein the first and second polypeptide chains of the binding protein comprise SEQ ID NOs:

74 and 73.

9. The binding protein of claim 8, wherein the binding protein is capable of:

(a) binding to VEGF with a dissociation constant (K_D) of at most about 7.0×

10<sup>−

10</sup> M, as measured by surface plasmon resonance, and/or blocking VEGF activity with an IC50 of at most about 3.8 nM, as measured in a VEGFR1 Competition ELISA; and

/or

(b) binding to DLL4 with a dissociation constant (K_D) of at most about 1.0×

10<sup>−

8</sup> M, as measured by surface plasmon resonance, and/or blocking DLL4 activity with an IC50 of at most about 1.09 nM, as measured in a Notch Competition ELISA.

10. A composition comprising the binding protein of claim 1 and at least one additional agent.

11. The composition of claim 10, wherein the at least one additional agent comprises one or more of a cytotoxic agent, a chemotherapeutic agent, an anti-angiogenic agent, an anti-CTLA4 antibody, a calcium channel blocker, an ACE inhibitor, FOLFIRI, paclitaxel, carboplatin, doxil, topotecan, and cisplatin.