IMMUNOGLOBULIN FUSION PROTEINS AND COMPOSITIONS THEREOF

• US 20160237156A1
• Filed: 07/11/2014
• Published: 08/18/2016
• Est. Priority Date: 07/11/2013
• Status: Abandoned Application

• Current Assignee: The Scripps Research Institute
• Original Assignee: The California Institute For Biomedical Research, The Scripps Research Institute
• Inventors: LIU, Tao, DU, Juanjuan, WANG, Ying, LIU, Yan, WANG, Feng, ZHANG, Yong, SCHULTZ, Peter G.
• Application Number:
  US14/903,489
• Publication Number:
  US 20160237156A1
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 47/65 Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
• CPC Class Codes: A61K 47/6889 Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• 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 1/06 Anti-spasmodics, e.g. drugs for colics, esophagic dyskinesia
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 25/04 Centrally acting analgesics, e.g. opioids
• 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 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 5/00 Drugs for disorders of the endocrine system
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/04 Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 14/43522 : from scorpions
• CPC Class Codes: C07K 14/47 : from mammals
• CPC Class Codes: C07K 14/505 : Erythropoietin [EPO]
• CPC Class Codes: C07K 14/535 : Granulocyte CSF; Granulocyte-macrophage CSF
• CPC Class Codes: C07K 14/57563 : Vasoactive intestinal peptide [VIP]; Related peptides
• CPC Class Codes: C07K 14/5759 : Products of obesity genes, e.g. leptin, obese (OB), tub, fat
• CPC Class Codes: C07K 14/605 : Glucagons
• CPC Class Codes: C07K 14/61 : Growth hormone [GH], i.e. somatotropin
• CPC Class Codes: C07K 14/64 : Relaxins
• CPC Class Codes: C07K 14/70503 : Immunoglobulin superfamily
• CPC Class Codes: C07K 14/811 : Serine protease (E.C. 3.4.21) inhibitors
• CPC Class Codes: C07K 16/1027 : Paramyxoviridae, e.g. respiratory syncytial virus
• 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 16/32 : against translation products of oncogenes
• 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 2317/565 : Complementarity determining region [CDR]
• 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 2318/10 : Immunoglobulin or domain(s) thereof as scaffolds for inserted non-Ig peptide sequences, e.g. for vaccination purposes
• CPC Class Codes: C07K 2319/00 : Fusion polypeptide
• CPC Class Codes: C07K 2319/30 : Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• 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 2319/50 : containing protease site
• CPC Class Codes: C07K 2319/55 : containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C07K 2319/73 : containing coiled-coiled motif (leucine zippers)

Abstract

Disclosed herein are immunoglobulin fusion proteins that have a first antibody region attached to an extender fusion region. The extender fusion region contains a therapeutic agent and a beta strand secondary structure. The extender fusion region may contain 7 or fewer consecutive amino acids based on or derived from an ultralong CDR3. Alternatively, the extender fusion region contains a rigid stalk protein structure, but does not contain an amino acid sequence based on or derived from an ultralong CDR3. The extender fusion region may also have one or more linkers or proteolytic cleavage sites. The immunoglobulin fusion proteins may have additional therapeutic agents and extender fusion regions. Also disclosed herein are pharmaceutical compositions of immunoglobulin fusion proteins and methods for using the immunoglobulin fusion proteins for the treatment or prevention of a disease or condition in a subject.

Claims

1-61. -61. (canceled)

62. An immunoglobulin fusion protein comprising:

a) a first antibody region; and

b) a first extender fusion region comprising a first therapeutic agent attached to a first extender peptide,

wherein the first extender peptide comprises a first beta strand secondary structure region having a first beta strand secondary structure;

wherein the first extender fusion region does not contain more than 7 consecutive amino acids from an ultralong complementary determining region 3 heavy chain of SEQ ID NO;

248.

63. The immunoglobulin fusion protein of claim 62, wherein the first extender fusion region comprises a second extender peptide comprising a second beta strand secondary structure region having a second beta strand secondary structure, and wherein the first beta strand secondary structure and second beta strand secondary structure form a beta sheet.

64. The immunoglobulin fusion protein of claim 62, wherein the first extender fusion region replaces a portion of a light chain of the first antibody region or a portion of the heavy chain of the first antibody region.

65. The immunoglobulin fusion protein of claim 62, wherein the first antibody region comprises an antibody selected from an anti-Her2 antibody, trastuzumab, an anti-CD47 antibody, palivizumab, and antigen binding fragments thereof.

66. The immunoglobulin fusion protein of claim 62, wherein the first antibody region comprises a heavy chain, wherein the heavy chain is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOS:

24-27, 29-33, 36-39, and 251-253, and a light chain, wherein the light chain is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOS;

21-23, 28, 34, 35, 40, 248-250 and 278.

67. The immunoglobulin fusion protein of claim 62, wherein the first beta strand secondary structure region comprises an amino acid sequence that is at least 90% homologous to a sequence selected from SEQ ID NOs:

109-128, 305-308.

68. The immunoglobulin fusion protein of claim 62, wherein the first beta strand secondary structure region comprises an amino acid sequence of ETKKYQX_nS (SEQ ID NO:

110), wherein n=1-8 and X is selected from a basic amino acid, an acidic amino acid, a polar amino acid, and a charged amino acid.

69. The immunoglobulin fusion protein of claim 63, wherein the first beta strand secondary structure region comprises an amino acid sequence of ETKKYQX_nS (SEQ ID NO:

306), wherein n=1-8 and X is selected from a basic amino acid, an acidic amino acid, a polar amino acid, and a charged amino acid, and wherein the second beta strand secondary structure region comprises an amino acid sequence of SX¹TX²NX³ (SEQ ID NO;

     128), wherein X¹, X², and X³ are independently selected from polar amino acids.

70. The immunoglobulin fusion protein of claim 63, wherein the second beta strand secondary structure region comprises an amino acid sequence of YX¹YX²Y, and wherein X¹ and X² are independently selected from polar amino acids.

71. The immunoglobulin fusion protein of claim 62, wherein the immunoglobulin fusion protein comprises an amino acid sequence that is at least about 90% identical to an amino acid sequence of any one of SEQ ID NOs:

76-108, 260-277, 298, 300, 302, and 304.

72. The immunoglobulin fusion protein of claim 62, wherein the immunoglobulin fusion protein further comprises a second therapeutic agent.

73. The immunoglobulin fusion protein of claim 72, wherein the immunoglobulin fusion protein further comprises a second antibody region, wherein the second antibody region comprises a second extender fusion region attached to the second antibody region, and wherein the second extender fusion region comprises the second therapeutic agent and an extender peptide comprising

a) an amino acid sequence having an alpha helical secondary structure, or

b) an amino acid sequence having a beta strand secondary structure, or

c) an amino acid sequence having no regular secondary structure.

74. The immunoglobulin fusion protein of claim 72, further comprising a second antibody region, wherein the second therapeutic agent is directly attached to the second antibody region.

75. The immunoglobulin fusion protein of claim 62, wherein first therapeutic agent is selected from bGCSF, hGCSF, bGMCSF, hGMCSF, GDF11, interferon-beta, interferon-alpha, interleukin 11 (IL-11), exendin-4, GLP-1, relaxin, oxyntomodulin, leptin, betatrophin, bovine growth hormone (bGH), human growth hormone (hGH), parathyroid hormone, erythropoietin, Moka1, VM-24, Mamba1, angiopoeitin-like 3 (ANGPTL3), CVX15, a CXCR4 ligand, a neutrophil elastase inhibitor, and homologs thereof.

76. The immunoglobulin fusion protein of claim 62, wherein the first therapeutic agent is represented by an amino acid sequence that is at least 90% homologous to a sequence selected from any one of SEQ ID NOs:

200-235 and/or encoded by a nucleic acid sequence that is at least 90% homologous to a sequences selected from SEQ ID NOs;

167-199.

77. The immunoglobulin fusion protein of claim 62, wherein the first antibody region comprises a BVK antibody or antigen binding fragment thereof and the first therapeutic agent comprises a Moka1 peptide, and wherein the first extender fusion region is grafted into a heavy chain of the BVK antibody.

78. The immunoglobulin fusion protein of claim 77, wherein the Moka1 peptide is represented by an amino acid sequence that is at least about 90% homologous to SEQ ID NO. 202.

79. The immunoglobulin fusion protein of claim 77, wherein the heavy chain of the BVK antibody is encoded by a nucleotide sequence that is at least 90% homologous to SEQ ID NO. 1.

80. The immunoglobulin fusion protein of claim 77, wherein the first beta strand secondary structure is represented by an amino acid sequence selected from SEQ ID NOS:

111, 119, and combinations thereof.

81. The immunoglobulin fusion protein of claim 77, wherein:

a) the BVK antibody comprises a heavy chain encoded by a nucleotide sequence of SEQ ID NO;

1, and a light chain encoded by a nucleotide sequence of SEQ ID NO;

14;

b) the Moka1 peptide is represented by an amino acid sequence of SEQ ID NO;

202;

c) the first beta strand secondary structure is represented by an amino acid sequence of SEQ ID NO;

111; and

d) a second beta strand secondary structure is represented by an amino acid sequence of SEQ ID NO;

119,

wherein the Moka1 peptide is grafted into the heavy chain of the BVK antibody.