Method for altering plasma retention and immunogenicity of antigen-binding molecule

Patent metadata

Field	Value
Publication number	US-10618965-B2
Application number	US-201214007947-A
Country	US
Kind code	B2
Filing date	Mar 30, 2012
Priority date	Feb 25, 2011
Publication date	Apr 14, 2020
Grant date	Apr 14, 2020

Abstract

The present invention demonstrated that the modification of the Fc region of an antigen-binding molecule into an Fc region that does not form in a neutral pH range a heterotetramer complex containing two molecules of FcRn and an active Fcγ receptor improved the pharmacokinetics of the antigen-binding molecule and reduced the immune response to the antigen-binding molecule. The present invention also revealed methods for producing antigen-binding molecules having the properties described above, and successfully demonstrated that pharmaceutical compositions containing as an active ingredient such an antigen-binding molecule or an antigen-binding molecule produced by a production method of the present invention have excellent features over conventional antigen-binding molecules in that when administered, they exhibit improved pharmacokinetics and reduced in vivo immune response.

First claim

The invention claimed is:

1. A method for reducing immunogenicity of an antibody, the method comprising:
identifying a first antibody comprising (a) an Fc region with a human FcRn-binding activity at pH 7.0 that is stronger than the human FcRn-binding activity at pH 7.0 of a native human IgG1 Fc region, and (b) an antigen-binding domain whose antigen-binding activity varies with pH as described in (i) below, or with calcium ion concentration as described in (ii) below:
(i) the antigen-binding activity is lower at pH 5.8 than at pH 7.4, wherein the ratio of the KD value for antigen-binding activity at pH 5.8 to the KD value for antigen-binding activity at pH 7.4 (KD (pH 5.8)/KD (pH 7.4)) is 2 or more when the KD (pH 5.8) and KD (pH 7.4) values are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM 2-(N-morpholino)ethanesulfonic acid (MES) pH 5.8 or pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, at 37° C.;
(ii) the antigen-binding activity is lower at a calcium ion concentration of 3 μM than at a calcium ion concentration of 2 mM, wherein the ratio of the KD value for antigen-binding activity at a calcium ion concentration of 3 μM to the KD value for antigen-binding activity at a calcium ion concentration of 2 mM (KD (3 μM)/ KD (2 mM)) is 2 or more when the KD (3 μM) and KD (2 mM) values are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM N-(2-acetamido)-2-aminoethanesulfonic acid (ACES) pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, and either 2 mM CaCl 2 or 3 μM CaCl 2 , at 37° C.;

producing a second antibody whose ability to form a heterocomplex with two molecules of the human FcRn and one molecule of an activating Fcγ receptor at pH 7.4 is reduced compared to the ability of the first antibody to form such a heterocomplex at pH 7.4, and whose ability to bind to the activating Fcγ receptor is decreased compared to the ability of the native human IgG1 Fc region to bind to the activating Fcγ receptor, the second antibody being identical to the first antibody except for one or more amino acids in the Fc region, wherein the activating Fcγ receptor is human FcγRIa, human FcγRIIa(R), human FcγRIIa(H), human FcγRIIIa(V), or human FcγRIIIa(F), and wherein at least one of the following positions in the Fc region of the second antibody is occupied by one of the amino acid residues listed for that position (by EU numbering):
Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Lys, Met, Phe, Pro, Ser, Thr, or Trp at position 234;
Ala, Asn, Asp, Gln, Glu, Gly, His, Ile, Lys, Met, Pro, Ser, Thr, Val, or Arg at position 235;
Arg, Asn, Gln, His, Leu, Lys, Met, Phe, Pro, or Tyr at position 236;
Ala, Asn, Asp, Gln, Glu, His, Ile, Leu, Lys, Met, Pro, Ser, Thr, Val, Tyr, or Arg at position 237;
Ala, Asn, Gln, Glu, Gly, His, Ile, Lys, Thr, Trp, or Arg at position 238;
Gln, His, Lys, Phe, Pro, Trp, Tyr, or Arg at position 239;
Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val at position 265;
Ala, Arg, Asn, Asp, Gln, Glu, Gly, His, Lys, Phe, Pro, Ser, Thr, Trp, or Tyr at position 266;
Arg, His, Lys, Phe, Pro, Trp, or Tyr at position 267;
Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 269;
Ala, Arg, Asn, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 270;
Arg, His, Phe, Ser, Thr, Trp, or Tyr at position 271;
Arg, Asn, Asp, Gly, His, Phe, Ser, Trp, or Tyr at position 295;
Arg, Gly, Lys, or Pro at position 296;
Ala at position 297;
Arg, Gly, Lys, Pro, Trp, or Tyr at position 298;
Arg, Lys, or Pro at position 300;
Lys or Pro at position 324;
Ala, Arg, Gly, His, Ile, Lys, Phe, Pro, Thr, Trp, Tyr, or Val at position 325;
Arg, Gln, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val at position 327;
Arg, Asn, Gly, His, Lys, or Pro at position 328;
Asn, Asp, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, Val, or Arg at position 329;
Pro or Ser at position 330;
Arg, Gly, or Lys at position 331;
Arg, Lys, or Pro at position 332; and
conducting an assay to confirm that the second antibody has decreased immunogenicity compared to the first antibody.

2. A method for reducing immunogenicity of an antibody, the method comprising:
identifying a first antibody comprising (a) an Fc region with a human FcRn-binding activity at pH 7.0 that is stronger than the human FcRn-binding activity at pH 7.0 of a native human IgG1 Fc region, and (b) an antigen-binding domain whose antigen-binding activity varies with pH as described in (i) below, or with calcium ion concentration as described in (ii) below:
(i) the antigen-binding activity is lower at pH 5.8 than at pH 7.4, wherein the ratio of the KD value for antigen-binding activity at pH 5.8 to the KD value for antigen-binding activity at pH 7.4 (KD (pH 5.8)/ KD (pH 7.4)) is 2 or more when the KD values for KD (pH 5.8) and KD (pH 7.4) are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM 2-(N-morpholino)ethanesulfonic acid (MES) at pH 5.8 or pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, at 37° C.;
(ii) the antigen-binding activity is lower at a calcium ion concentration of 3 μM than at a calcium ion concentration of 2 mM, wherein the ratio of the KD value for antigen-binding activity at a calcium ion concentration of 3 μM to the KD value for antigen-binding activity at a calcium ion concentration of 2 mM (KD (3 μM) / KD (2 mM)) is 2 or more when the KD values for KD (3 μM) and KD (2 mM) are determined using a surface plasmon resonance technique in which the antibody is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM N-(2-acetamido)-2-aminoethanesulfonic acid (ACES) at pH 7.4, 150 mM NaCl, 0.05% polysorbate 20, and either 2mM CaCl 2 or 3 μM CaCl 2 , at 37° C.;

producing a second antibody whose ability to form a heterocomplex with two molecules of the human FcRn and one molecule of an activating Fcγ receptor at pH 7.4 is reduced compared to the ability of the first antibody to form such a heterocomplex at pH 7.4, the second antibody being identical to the first antibody except at one or more positions in the Fc region, including
(a) position 238 (EU numbering), which in the second antibody is Asp and in the first antibody is not Asp, or
(b) position 328 (EU numbering), which in the second antibody is Glu and in the first antibody is not Glu,

wherein the second antibody binds more strongly to human FcγRIIb than to the activating Fcγ receptor, wherein the activating Fcγ receptor is human FcγRIa, human FcγRIIa(R), human FcγRIIa(H), human FcγRIIIa(V), or human FcγRIIIa(F); and
conducting an assay to confirm that the second antibody has decreased immunogenicity compared to the first antibody.

3. A method for reducing immunogenicity of an antibody, the method comprising:
identifying a first antibody comprising
(a) a first Fc region that binds to a human FcRn at pH 7.0 more strongly than a native human IgG1 Fc region binds to the human FcRn at pH 7.0, and that is able to form a heterocomplex with two molecules of the human FcRn and one molecule of an activating Fcγ receptor at pH 7.4, and
(b) an antigen-binding domain whose antigen-binding activity varies with pH as described in (i) below, or with calcium ion concentration as described in (ii) below:
(i) the antigen-binding activity is lower at pH 5.8 than at pH 7.4, wherein the ratio of the KD value for antigen-binding activity at pH 5.8 to the KD value for antigen-binding activity at pH 7.4 (KD (pH 5.8)/ KD (pH 7.4)) is 2 or more when the KD (

Assignees

• Igawa Tomoyuki
• Maeda Atsuhiko
• Haraya Kenta
• Iwayanagi Yuki
• Tachibana Tatsuhiko
• Mimoto Futa
• Kuramochi Taichi
• Katada Hitoshi
• Kadono Shojiro
• Chugai Pharmaceutical Co Ltd

Inventors

• Igawa Tomoyuki
• Maeda Atsuhiko
• Haraya Kenta
• Iwayanagi Yuki
• Tachibana Tatsuhiko
• Mimoto Futa
• Kuramochi Taichi
• Katada Hitoshi
• Kadono Shojiro

Classifications

• A61P5/18

  of the parathyroid hormones · CPC title

• A61P1/04

  for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants · CPC title

• A61P25/00

  Drugs for disorders of the nervous system · CPC title

• A61P7/00

  Drugs for disorders of the blood or the extracellular fluid · CPC title

• A61P7/06

  Antianaemics · CPC title