ULTRALONG COMPLEMENTARITY DETERMINING REGIONS AND USES THEREOF

• US 20140086871A1
• Filed: 01/09/2013
• Published: 03/27/2014
• Est. Priority Date: 01/09/2012
• Status: Active Grant

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Smider, Vaughn, Schultz, Peter, Wilson, Ian, Bazirgan, Omar A., Ekiert, Dan, Mao, Hongyuan Helen, Wang, Feng, Zhang, Yong
• Granted Patent: US 10,774,132 B2
• Time in Patent Office: Days
• US Class Current: 424/85.6
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/53 Colony-stimulating factor [CSF]
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/10 from RNA viruses
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 2317/20 characterized by taxonomic origin
• CPC Class Codes: C07K 2317/50 characterized by immunoglobulin fragments
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/569 Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2318/00 Antibody mimetics or scaffolds
• 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 2319/50 containing protease site
• CPC Class Codes: C07K 2319/55 containing a fusion with a toxin, e.g. diphteria toxin
• CPC Class Codes: C12N 15/09 Recombinant DNA-technology
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/63 : Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
• CPC Class Codes: C12N 5/00 : Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00

Abstract

Disclosed herein are immunoglobulin constructs comprising at least one immunoglobulin domain or fragment thereof; and a therapeutic polypeptide or derivative or variant thereof attached to or inserted into said immunoglobulin domain. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a knob domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said knob domain of the CDR3H. Also provided are immunoglobulin constructs comprising a mammalian immunoglobulin heavy chain comprising at least a portion of a stalk domain in the complementarity-determining region 3 (CDR3H) or fragment thereof; and a therapeutic polypeptide attached to or inserted into said stalk domain of the CDR3H. Also described herein are methods and compositions comprising the immunoglobulin constructs described herein for treatment and prevention of a disease or condition in a subject.

Claims

1-47. -47. (canceled)

48. A recombinant antibody or fragment thereof, wherein at least a portion of the recombinant antibody or fragment thereof is based on or derived from at least a portion of an ultralong CDR3.

50. The antibody of claim 48, wherein the antibody is a chimeric, human engineered, or humanized antibody.

51. The antibody of claim 48, wherein the antibody is a bovine, bovinized, bovine engineered, or fully bovine antibody.

54. The antibody of claim 48 further comprising a non-antibody sequence.

60. The antibody of claim 54, wherein the non-antibody sequence is a derived from a growth factor, cytokine, hormone, or toxin.

61. The antibody of claim 60, wherein the growth factor is a GCSF, GMCSF or FGF21.

62. The antibody of claim 60, wherein the GCSF is a bovine GCSF or human GCSF.

63. The antibody of claim 54, wherein the non-antibody sequence is a beta-interferon, exendin-4, GLP-1, somatostatin, erythropoietin, Moka1, VM-24, or protoxin2.

69. The antibody of claim 54, wherein the non-antibody sequence is 50% or more homologous to a sequence selected from the group comprising SEQ ID NOS:

323-332.

70. The antibody of claim 48, wherein the ultralong CDR3 is based on or derived from a cow ultralong CDR3.

77. The antibody of claim 48, wherein the antibody comprises one or more amino acid residues based on or derived from a stalk domain of the ultralong CDR3, a knob domain of the ultralong CDR3, or a combination thereof.

82. The antibody of claim 48, wherein the antibody comprises a sequence that is 50% or more homologous to a sequence selected from any one of SEQ ID NOS:

157-224 and 235-295.

86. The antibody of claim 48, wherein the portion of the ultralong CDR3 is based on or derived from a BLV1H12 ultralong CDR3 sequence.

87. The antibody of claim 48, wherein the portion of the ultralong CDR3 is based on or derived from a sequence that is 50% or more homologous to a BLV1H12 ultralong CDR3 sequence.

89. The antibody of claim 48, wherein the portion of the ultralong CDR3 is based on or derived from a sequence that is 50% or more homologous to a BLV5B8, BLVCV1, BLV5D3, BLV8C11, BF1H1, or F18 ultralong CDR3 sequence.

125. The antibody of claim 48, wherein the antibody or binding fragment thereof comprises a sequence selected from SEQ ID NOS:

24-37, the antibody or binding fragment thereof encoded by the DNA of SEQ ID NOS;

2-15.

126. The antibody of claim 48, wherein the antibody or binding fragment thereof comprises a sequence that is 50% or more homologous to a sequence selected from SEQ ID NO:

23, the antibody or binding fragment thereof encoded by a DNA sequence that is 50% or more homologous to a sequence of SEQ ID NO;

1.

131. The antibody of claim 48, further comprising one or more linker sequences.

139. The antibody of claim 131, wherein the one or more linker sequences comprise (a) the sequence (GGGGS)_n and wherein n=1 to 5;

(b) the sequence GGGSGGGGS;

(c) the sequence GGGGSGGGS;

or (d) a combination of (a)-(c).

149. The antibody of claim 54 further comprising one or more cleavage sites.

150. The antibody of claim 149, wherein the one or more cleavage sites comprise a recognition site for a protease.

153. The antibody of claim 149, wherein the one or more cleavage sites are between the antibody sequence and the non-antibody sequence.

166. A polynucleotide comprising a nucleic acid sequence that encodes the antibody of claim 48.

167. A vector comprising a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that encodes the antibody of claim 48.

168. A host cell comprising a polynucleotide, wherein the polynucleotide comprises a nucleic acid sequence that encodes the antibody of claim 48.

169. A method of producing an antibody or binding fragment thereof comprising an ultralong CDR3 or fragment thereof comprising culturing the host cell of claim 168 under conditions wherein the polynucleotide sequence is expressed and the antibody or binding fragment thereof comprising an ultralong CDR3 or fragment thereof is produced.

171. A pharmaceutical composition comprising the antibody of claim 48.

173. The pharmaceutical composition of claim 171, further comprising a therapeutic agent, a pharmaceutically acceptable excipient, or a combination thereof.

175. A method of treating a disease or condition in a subject in need thereof comprising administering to the mammal a therapeutically effective amount of the antibody of claim 48.

176. The method of claim 175, wherein the antibody comprises an ultralong CDR3 sequence and a non-antibody sequence.

177. The method of claim 175, wherein the non-antibody sequence is selected from the group comprising Moka1, Vm24, GLP-1, Exendin-4, beta-interferon, human EPO, human FGF21, human GMCSF, human interferon-beta, bovine GCSF, human GCSF, protoxin2 and a derivative or variant thereof.

184. The method of claim 175, wherein the disease or condition is mastitis.

186. The method of claim 175, wherein the mammal is a bovine or human.

189. The antibody of claim 149, wherein the one or more cleavage sites are at the N-terminus of the non-antibody sequence, at the C-terminus of the non-antibody sequence, or a combination thereof.

49. (canceled)

52. (canceled)

53. (canceled)

55-59. -59. (canceled)

64-68. -68. (canceled)

71-76. -76. (canceled)

78-81. -81. (canceled)

83-85. -85. (canceled)

88. (canceled)

90-124. -124. (canceled)

127-130. -130. (canceled)

132-138. -138. (canceled)

140-148. -148. (canceled)

151-152. -152. (canceled)

154-165. -165. (canceled)

170. (canceled)

172. (canceled)

174. (canceled)

178-183. -183. (canceled)

185. (canceled)

187-188. -188. (canceled)