Methods for producing antibodies promoting disappearance of antigens having plurality of biological activities

Patent metadata

Field	Value
Publication number	US-11827699-B2
Application number	US-201816028140-A
Country	US
Kind code	B2
Filing date	Jul 5, 2018
Priority date	Sep 30, 2011
Publication date	Nov 28, 2023
Grant date	Nov 28, 2023

Abstract

The present inventors newly discovered that even if an antigen-binding molecule inhibits in vitro some of the physiological activities of an antigen having two or more physiological activities without inhibiting the remaining physiological activities, the molecule can promote elimination of the antigen from blood (from serum or plasma) and as a result reduce the physiological activities in vivo, when the antigen-binding molecule is conferred with the properties: (i) of binding to human FcRn under an acidic pH range condition; (ii) of binding under a neutral pH range condition to human Fc receptor stronger than native human IgG, and (iii) that its antigen-binding activity alters according to the ion concentration.

First claim

The invention claimed is:

1. A method for producing an antibody comprising an antigen binding domain and an Fc region, the method comprising:
(a) selecting an antigen comprising (i) a first target molecule binding domain that, when bound to a first target molecule, produces a first physiological activity, and (ii) a second target molecule binding domain that, when bound to a second target molecule, produces a second physiological activity, wherein the first and second target molecule binding domains are at different locations on the antigen, wherein the first and second target molecules are different, wherein the first and second physiological activities are different, and wherein the antigen cannot bind to both of the target molecules simultaneously;
(b) identifying one or more antigen-binding domains that bind to the antigen and, when bound to the antigen, inhibit the antigen's ability to bind to the first target molecule but do not inhibit the antigen's ability to bind to the second target molecule;
(c) selecting, from the one or more antigen-binding domains of (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, or with both:
(i) the antigen-binding activity is lower at pH 6.0 than at pH 7.4, wherein the ratio of the KD value for antigen binding at pH 6.0 to the KD value for antigen binding at pH 7.4 (KD (pH 6)/KD (pH 7.4)) is 2 or more when KD values are determined using a surface plasmon resonance technique in which a polypeptide comprising the antigen-binding domain is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM N-2-aminoethanesulfonic acid (ACES), 150 mM NaCl, 0.05% polysorbate 20, 37° C., and either pH 6.0 or pH 7.4;
(ii) the antigen-binding activity is lower at a calcium ion concentration of 3 μM than at a second calcium ion concentration of 2 mM, wherein the ratio of the KD value for antigen binding at a calcium ion concentration of 3 μM to the KD value for antigen binding at a calcium ion concentration of 2 mM (KD (3 μM Ca++)/KD (2 mM Ca++)) is 2 or more when KD values are determined using a surface plasmon resonance technique in which a polypeptide comprising the antigen-binding domain is immobilized, the antigen serves as analyte, and the following conditions are used: 10 mM ACES, 150 mM NaCl, 0.05% (w/v) polysorbate 20, 37° C., pH 7.4, and either 3 μM CaCl 2 ) or 2 mM CaCl 2 );

(d) identifying an IgG Fc region that
(i) differs from the Fc region of a native human IgG by amino acid substitution, insertion, or deletion at one or more positions,
(ii) binds to a human neonatal Fc receptor (FcRn) at pH 6, and
(iii) binds, at pH 7.4, to a human Fc receptor with greater affinity than the affinity with which the native human IgG binds to the human Fc receptor, wherein the native human IgG is of the same isotype as the identified IgG Fc region, and the native human IgG is a native human IgG1, a native human IgG2, a native human IgG3, or a native human IgG4; and

(e) preparing an antibody comprising the antigen-binding domain selected in (c) and the IgG Fc region identified in (d).

2. The method of claim 1 , wherein the amino acid sequence of the identified IgG Fc region is not identical to the amino acid sequence of any naturally occurring IgG Fc region.

3. The method of claim 1 , wherein (e) comprises expressing DNA encoding the antibody, thereby producing the antibody.

4. The method of claim 1 , wherein the human Fc receptor of (d)(iii) is the human FcRn.

5. The method of claim 4 , wherein the human Fc receptor of (d)(iii) is a human Fcγ receptor.

6. The method of claim 1 , wherein introduction of the antibody into an individual whose plasma comprises the antigen reduces the concentration of the antigen in the plasma of the individual.

7. The method of claim 1 , wherein the antigen-binding domain selected in (c) comprises at least one histidine residue.

8. The method of claim 4 , wherein the one or more positions are selected from the following positions (all by EU numbering): 234, 235, 236, 237, 238, 239, 244, 245, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 260, 262, 265, 267, 270, 272, 274, 279, 280, 282, 283, 284, 285, 286, 288, 289, 293, 295, 297, 298, 303, 305, 307, 308, 309, 311, 312, 313, 314, 315, 316, 317, 318, 325, 326, 327, 328, 329, 330, 332, 334, 338, 339, 340, 341, 343, 345, 360, 361, 362, 375, 376, 377, 378, 380, 382, 384, 385, 386, 387, 389, 390, 391, 413, 422, 423, 424, 427, 428, 430, 431, 433, 434, 435, 436, 437, 438, 440, and 442.

9. The method of claim 8 , wherein the one or more positions include at least one from the following positions (all by EU numbering), substituted with the indicated amino acid:
position 234: Arg;
position 235: Gly, Lys, or Arg;
position 236: Ala, Asp, Lys, or Arg;
position 237: Lys, Met, or Arg;
position 238: Ala, Asp, Lys, Leu, or Arg;
position 239: Asp or Lys;
position 244: Leu;
position 245: Arg;
position 248: Ile or Tyr;
position 249: Pro;
position 250: Ala, Glu, Phe, Ile, Met, Gln, Ser, Val, Trp, Gly, His, Leu, Asn, or Tyr;
position 251: Arg, Asp, Glu, or Leu;
position 252: Phe, Ser, Thr, Trp, or Tyr;
position 253: Val,
position 254: Ala, Gly, His, Le, Gin, Ser, Val, or Thr;
position 255: Ala, Asp, Phe, His, le, Lys, Leu, Met, Asn, Gin, Arg, Gly, Ser, Trp, Tyr, or Glu;
position 256: Ala, Asp, Glu, Arg, Asn, Pro, Thr, Ser, or Gin;
position 257: Ala, Gly, Ile, Leu, Met, Asn, Ser, Thr, or Val;
position 258: Asp or His;
position 260: Ser;
position 262: Leu;
position 265: Ala;
position 267: Met or Leu;
position 270: Lys or Phe;
position 272: Ala, Leu, or Arg;
position 274: Ala;
position 279: Leu, Ala, Asp, Gly, His, Met, Asn, Gin, Arg, Ser, Thr, Trp, or Tyr;
position 280: Ala, Gly, His, Lys, Asn, Gin, Arg, Ser, Thr, or Glu;
position 282: Ala or Asp;
position 283: Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gin, Arg, Ser, Thr, Trp, or Tyr;
position 284: Lys;
position 285: Asn;
position 286: Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gin, Arg, Ser, Thr, Val, Trp, Tyr, or Glu;
position 288: Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gin, Arg, Val, Trp, Tyr, or Ser;
position 289: His;
position 293: Val;
position 295: Met;
position 297: Ala;
position 298: Gly;
position 303: Ala;
position 305: Ala or Thr;
position 307: Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gin, Arg, Ser, Val, Trp, or Tyr;
position 308: Ala, Phe, lie, Leu, Met, Pro, Gin, or Thr;
position 309: Ala, Asp, Glu, Pro, His, or Arg;
position 311: Ala, His, Glu, Lys, Leu, Met, Ser, Val, Trp, or Ile;
position 312: Ala, Asp, Pro, or His;
position 313: Tyr or Phe;
position 314: Ala, Leu, Lys, or Arg;
position 315: Ala, Asp, Glu, Phe, Gly, lie, Lys, Leu, Met, Gin, Arg, Ser, Thr, Val, Trp, Tyr, or His;
position 316: Ala, Glu, Phe, His, lie, Lys, Leu, Met, Asn, Pro, Gin, Arg, Ser, Thr, Val, Trp, or Asp;
position 317: Ala or Pro;
position 318: Asn or Thr;
position 325: Ala, Gly, Met, Leu, Ile, or Ser;
position 326: Asp;
position 327: Gly;
position 328: Arg, Asp, Glu, or Tyr;
position 329: Lys or Arg;
position 330: Leu;
position 332: Glu, Phe, His, Lys, Leu, Met, Arg, Ser, Trp, or Val;
position 334: Leu;
position 338: Ala;
position 339: Asn, Thr, or Trp;
position 340: Ala;
position 341: Pro;
position 343: Glu, His, Lys, Gin, Arg, Thr, or Tyr;
position 345: Ala;
position 360: His;
position 361: Ala;
position 362: Ala;
position 375: Ala or Arg;
position 376: Ala, Gly, le, Met, Pro, Thr, or Val;
position 377: Lys;
position 378: Asp, Asn, or Val;
position 380: Ala, Asn, Thr, or Ser;
position 382:

Assignees

• Chugai Pharmaceutical Co Ltd

Inventors

• Igawa Tomoyuki
• Maeda Atsuhiko
• Tamba Shigero
• Kitazawa Takehisa
• Baba Takeshi
• Ruike Yoshinao
• Nezu Junichi

Classifications

• C07K16/24 Primary

  against cytokines, lymphokines or interferons · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• C07K16/18 Primary

  against material from animals or humans · CPC title

• C07K16/22

  against growth factors {; against growth regulators} · CPC title

• C07K16/248

  IL-6 · CPC title