CYTOTOXICITY-INDUCING THERAPEUTIC AGENT

• US 20140112914A1
• Filed: 11/30/2011
• Published: 04/24/2014
• Est. Priority Date: 11/30/2010
• Status: Active Grant

• Current Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Original Assignee: Chugai Seiyaku Kabushiki Kaisha (Roche Holding AG)
• Inventors: Ishiguro, Takahiro, Sakamoto, Akihisa, Igawa, Tomoyuki, Kuramochi, Taichi, Nezu, Junichi, Narita, Atsushi, Kawai, Yumiko
• Granted Patent: US 11,066,483 B2
• Time in Patent Office: Days
• US Class Current: 424/133.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/2809 against the T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/303 Liver or Pancreas
• 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 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/41 Glycosylation, sialylation, or fucosylation
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• 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 2317/60 characterized by non-natural combinations of immunoglobulin fragments
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/73 : Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/94 : Stability, e.g. half-life, pH, temperature or enzyme-resistance
• 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

Abstract

By replacing the antigen-binding domain, the present inventors discovered novel polypeptide complexes that retain BiTE'"'"'s strong anti-tumor activity and excellent safety properties, as well as have long half-life in blood and can damage various different target cells.

Claims

1-68. -68. (canceled)

69. A polypeptide complex that comprises:

(1) an antigen-binding domain;

(2) an Fc domain that has reduced Fcγ

receptor-binding activity compared to a wild-type Fc region of IgG1; and

(3) a T cell receptor complex-binding domain.

70. The polypeptide complex of claim 69, wherein the T cell receptor complex-binding domain is a T cell receptor-binding domain.

71. The polypeptide complex of claim 69, wherein the T cell receptor complex-binding domain is a CD3-binding domain.

72. The polypeptide complex of claim 69, wherein the antigen-binding domain is a bivalent antigen-binding domain.

73. The polypeptide complex of claim 72, wherein the bivalent antigen-binding domain is a domain having an F(ab′

)2 structure.

74. The polypeptide complex of claim 73, wherein the F(ab′

)2 structure comprises two polypeptides, each comprising a portion of a heavy chain constant region;

the Fc domain comprises two polypeptides; and

each portion of a heavy chain constant region is linked to one of the two Fc domain polypeptides.

75. The polypeptide complex of claim 74, wherein the T cell receptor complex-binding domain is a CD3-binding domain;

the Fc domain comprises two CH3 domains; and

the CD3-binding domain is linked to one or both of the CH3 domains.

76. The polypeptide complex of claim 75, wherein the CD3-binding domain comprises a heavy chain Fv fragment and a light chain Fv fragment;

the heavy chain Fv fragment of the CD3-binding domain is linked to one of the two CH3 domains present in the Fc domain; and

the light chain Fv fragment of the CD3-binding domain is linked to the second CH3 domain.

77. The polypeptide complex of claim 76, further comprising a CH1 domain linked to the heavy chain Fv fragment of the CD3-binding domain and an antibody light chain constant domain (CL) linked to the light chain Fv fragment of the CD3-binding domain.

78. The polypeptide complex of claim 74, wherein the T cell receptor complex-binding domain is a CD3-binding domain;

the F(ab′

)2 structure comprises two CL domains; and

the CD3-binding domain is linked to one or both of the two CL domains of the F(ab′

)2 structure.

79. The polypeptide complex of claim 74, wherein the T cell receptor complex-binding domain is a CD3-binding domain;

the F(ab′

)2 structure comprises two VHs; and

the CD3-binding domain is linked to one or both of the two VHs of the F(ab′

)2 structure.

80. The polypeptide complex of claim 74, wherein the T cell receptor complex-binding domain is a CD3-binding domain;

the F(ab′

)2 structure comprises two VLs; and

the CD3-binding domain is linked to one or both of the two VLs of the F(ab′

)2 structure.

81. The polypeptide complex of claim 71, wherein the CD3-binding domain is a Fv.

82. The polypeptide complex of claim 71, wherein the CD3-binding domain is a Fab.

83. The polypeptide complex of claim 71, wherein the CD3-binding domain is a scFv.

84. The polypeptide complex of claim 71, wherein the CD3-binding domain is monovalent.

85. The polypeptide complex of claim 69, wherein the antigen-binding domain comprises a monovalent scFv and a monovalent Fab.

86. The polypeptide complex of claim 85, wherein

the Fc domain comprises two polypeptides;

the T-cell receptor complex-binding domain is a CD3-binding scFv;

the monovalent scFv is linked to one of the polypeptides of the Fc domain via the CD3-binding scFv;

the monovalent Fab comprises (a) a heavy chain Fv fragment linked to a CH1 domain, and (b) a light chain Fv fragment linked to a CL domain; and

the heavy chain Fv fragment of the monovalent Fab is linked to the second polypeptide of the Fc domain via the CH1 domain.

87. The polypeptide complex of claim 69, wherein the antigen-binding domain comprises two monovalent scFv.

88. The polypeptide complex of claim 87, wherein

the Fc domain comprises two polypeptides;

the T-cell receptor complex-binding domain is a CD3-binding domain comprising a heavy chain Fv fragment and a light chain Fv fragment;

one of the monovalent scFv of the antigen-binding domain is linked to one of the polypeptides of the Fc domain via the heavy chain Fv fragment of the CD3-binding domain; and

the second monovalent scFv of the antigen-binding domain is linked to the second polypeptide of the Fc domain via the light chain Fv fragment of the CD3-binding domain.

89. The polypeptide complex of claim 87, wherein

the Fc domain comprises two polypeptides;

the T-cell receptor complex-binding domain is a CD3-binding scFv;

one of the monovalent scFv of the antigen-binding domain is linked to one of the polypeptides of the Fc domain via the CD3-binding scFv; and

the second monovalent scFv of the antigen-binding domain is linked to the second polypeptide of the Fc domain.

90. The polypeptide complex of claim 69, wherein the antigen-binding domain and the T cell receptor complex-binding domain each is a monovalent Fab.

91. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a heavy chain Fv fragment linked to one of the polypeptides of the Fc domain via a CH1 domain, and (b) a light chain Fv fragment linked to a first CL domain; and

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to the second polypeptide of the Fc domain via a second CH1 domain, and (b) a light chain Fv fragment linked to a second CL domain.

92. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a heavy chain Fv fragment linked to one of the polypeptides of the Fc domain via a CH1 domain, and (b) a light chain Fv fragment linked to a first CL domain; and

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to a second CH1 domain, and (b) a light chain Fv fragment linked to the second polypeptide of the Fc domain via a second CL domain.

93. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a heavy chain Fv fragment linked to one of the polypeptides of the Fc domain via a first CH1 domain, and (b) a light chain Fv fragment linked to a first CL domain; and

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to the second polypeptide of the Fc domain via a second CL domain, and (b) a light chain Fv fragment linked to a second CH1 domain.

94. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a light chain Fv fragment linked to one of the polypeptides of the Fc domain via a first CH1 domain, and (b) a heavy chain Fv fragment linked to a first CL domain; and

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to the second polypeptide of the Fc domain via a second CH1 domain, and (b) a light chain Fv fragment linked to a second CL domain.

95. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a heavy chain Fv fragment linked to one of the polypeptides of the Fc domain via a first CL domain, and (b) a light chain Fv fragment linked to a first CH1 domain; and

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to the second polypeptide of the Fc domain via a second CH1 domain, and (b) a light chain Fv fragment linked to a second CL domain.

96. The polypeptide complex of claim 90, wherein

the Fc domain comprises two polypeptides;

the monovalent Fab forming the antigen-binding domain comprises (a) a heavy chain Fv fragment linked to one of the polypeptides of the Fc domain via a first CH1 domain, and (b) a light chain Fv fragment linked to a first CL domain;

the monovalent Fab forming the T cell receptor complex-binding domain comprises (a) a heavy chain Fv fragment linked to the second polypeptide of the Fc domain via a second CH1 domain, and (b) a light chain Fv fragment linked to a second CL domain; and

electric charges of the CH1 and CL domains are controlled so that the heavy chain Fv fragment of the monovalent Fab forming the antigen-binding domain assembles with the light chain Fv fragment of the monovalent Fab forming the antigen-binding domain, and/or the heavy chain Fv fragment of the monovalent Fab forming the T cell receptor complex-binding domain assembles with the light chain Fv fragment of the monovalent Fab forming the T cell receptor complex-binding domain.

97. The polypeptide complex of claim 96, wherein an amino acid residue in the second CH1 domain has the same type of electric charge as an amino acid residue in the first CL domain.

98. The polypeptide complex of claim 96, wherein an amino acid residue in the first CH1 domain has the same type of electric charge as an amino acid residue in the second CL domain.

99. The polypeptide complex of claim 96, wherein an amino acid residue in the second CH1 domain has the same type of electric charge as an amino acid residue in the first CL domain, and an amino acid residue in the first CH1 domain has the same type of electric charge as an amino acid residue in the second CL domain.

100. The polypeptide complex of claim 97, wherein an amino acid residue in the second CH1 domain has an electric charge opposite to that of an amino acid residue in the second CL domain.

101. The polypeptide complex of claim 98, wherein an amino acid residue in the first CH1 domain has an electric charge opposite to that of an amino acid residue in the first CL domain.

102. The polypeptide complex of claim 90, wherein the T cell receptor complex-binding domain is a T cell receptor-binding domain.

103. The polypeptide complex of claim 102, wherein the T cell receptor-binding domain is a CD3-binding domain.

104. The polypeptide complex of claim 100, wherein the amino acid residues with opposite electric charges are at positions (EU numbering) within the second CH1 domain and the first CL domain defined by one or more of the following combinations:

(a) position 147 in the CH1 domain and position 180 in the CL domain;

(b) position 147 in the CH1 domain and position 131 in the CL domain;

(c) position 147 in the CH1 domain and position 164 in the CL domain;

(d) position 147 in the CH1 domain and position 138 in the CL domain;

(e) position 147 in the CH1 domain and position 123 in the CL domain;

(f) position 175 in the CH1 domain and position 160 in the CL domain; and

(g) position 213 in the CH1 domain and position 123 in the CL domain.

105. The polypeptide complex of claim 104, wherein the amino acid residues at position 213 in the second CH1 domain and position 123 in the first CL domain (EU numbering) have opposite electric charges.

106. The polypeptide complex of claim 104, wherein each of the combinations of two amino acid residues having opposite electric charges includes one residue from group (i) and one from group (ii):

(i) glutamic acid (E) and aspartic acid (D); and

(ii) lysine (K), arginine (R), and histidine (H).

107. The polypeptide complex of claim 104, wherein the amino acid residues having opposite electric charges include Lys at positions 147 and 175 (EU numbering) in the second CH1 domain, and Glu at one or more of positions 131, 160, and 180 (EU numbering) in the first CL domain.

108. The polypeptide complex of claim 104, wherein the amino acid residues having opposite electric charges include Glu at positions 147 and 175 (EU numbering) in the second CH1 domain, and Lys at positions 131, 160, and 180 (EU numbering) in the first CL domain.

109. The polypeptide complex of claim 108, wherein the amino acid at position 213 (EU numbering) in the second CH1 domain is Glu, and the amino acid at position 123 (EU numbering) in the first CL domain is Lys.

110. The polypeptide complex of claim 69, wherein the Fc domain exhibits impaired binding to an Fcγ

receptor selected from Fcγ

I, Fcγ

IIA, Fcγ

IIB, Fcγ

IIIA, and Fcγ

IIIB, compared to binding to the selected Fcγ

receptor exhibited by wild-type IgG1 Fc region.

111. The polypeptide complex of claim 69, wherein the Fc domain comprises a sequence corresponding to the Fc region of any one of SEQ ID NOs:

23, 24, 25, and 26, with an amino acid substitution.

112. The polypeptide complex of claim 111, wherein

the amino acid sequence from positions 118 to 260 (EU numbering) of the Fc domain is the sequence from the corresponding portion of the Fc region contained in SEQ ID NO:

24;

or

the amino acid sequence at positions 261 to 447 (EU numbering) of the Fc domain is the sequence from the corresponding portion of the Fc region contained in SEQ ID NO;

26.

113. The polypeptide complex of claim 111, wherein the substitution is at any one of the following positions:

220, 226, 229, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 264, 265, 266, 267, 269, 270, 295, 296, 297, 298, 299, 300, 325, 327, 328, 329, 330, 331, and 332 (EU numbering).

114. The polypeptide complex of claim 113, wherein the Fc domain comprises a sequence corresponding to the Fc region of any one of SEQ ID NO:

23, with an amino acid substitution.

115. The polypeptide complex of claim 114, wherein the substitution is at position 233, 234, 235, 236, 237, 327, 330, or 331 (EU numbering), the substitution being by an amino acid from a corresponding position in IgG2 or IgG4.

116. The polypeptide complex of claim 114, wherein the substitution is at position 234, 235, or 297 (EU numbering).

117. The polypeptide complex of claim 116, wherein the substitution is with alanine.

118. The polypeptide complex of claim 111, wherein the Fc domain comprises two polypeptides that have different sequences.

119. The polypeptide complex of claim 118, wherein

in one of the two polypeptides forming the Fc domain, the amino acids at position 349 and 366 (EU numbering) are substituted with cysteine and tryptophan, respectively;

and

in the second polypeptide forming the Fc domain, the amino acids at positions 356, 366, 368, and 407 (EU numbering) are substituted with cysteine, serine, alanine, and valine, respectively.

120. The polypeptide complex of claim 118, wherein:

in one of the two polypeptides forming the Fc domain, the amino acid at position 356 is substituted with lysine;

in the second of the two polypeptides forming the Fc domain, the amino acid at position 439 is substituted with glutamic acid; and

in either the first or the second of the two polypeptides forming the Fc domain, the amino acid at position 435 is substituted with arginine (all positions being by EU numbering).

121. The polypeptide complex of claim 119, wherein the sequence glycine-lysine is deleted from the carboxyl termini of both the first and the second of the two polypeptides forming the Fc domain.

122. The polypeptide complex of claim 72, wherein the bivalent antigen-binding domain binds to two copies of the same epitope.

123. The polypeptide complex of claim 122, wherein the epitope is present in a protein consisting of the amino acid sequence of SEQ ID NO:

2.

124. The polypeptide complex of claim 122, wherein the epitope is present in a protein consisting of the amino acid sequence of SEQ ID NO:

4.

125. The polypeptide complex of claim 72, wherein the bivalent antigen-binding domain binds to two different epitopes.

126. The polypeptide complex of claim 125, wherein at least one of the two different epitopes is present in a protein consisting of the amino acid sequence of SEQ ID NO:

2.

127. The polypeptide complex of claim 125, wherein at least one of the two different epitopes is present in a protein consisting of the amino acid sequence of SEQ ID NO:

4.

128. A set of one to four polynucleotides that together encode all polypeptide chains that make up the polypeptide complex of claim 69.

129. A set of one or more vectors that together comprise the set of one to four polynucleotides of claim 128.

130. A cell comprising the set of one or more vectors of claim 129.

131. A method for producing a polypeptide complex, the method comprising culturing the cell of claim 130 under conditions sufficient to produce the polypeptide complex, and isolating the polypeptide complex from the culture supernatant.

132. A pharmaceutical composition for inducing cellular cytotoxicity, wherein the pharmaceutical composition comprises as an active ingredient the polypeptide complex of claim 69.

133. A method for treating cancer in a subject, the method comprising administering to a subject in need thereof the polypeptide complex of claim 69.

134. The method of claim 133, wherein the cancer is liver cancer or lung cancer.