Optimized anti-CD20 monoclonal antibodies having Fc variants

• US 20060121032A1
• Filed: 06/30/2005
• Published: 06/08/2006
• Est. Priority Date: 03/03/2003
• Status: Active Grant

• Current Assignee: Xencor, Inc.
• Original Assignee: Xencor, Inc.
• Inventors: Lazar, Gregory Alan, Vafa, Omid, Dahiyat, Bassil I.
• Granted Patent: US 8,084,582 B2
• Time in Patent Office: Days
• US Class Current: 424/144.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• 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 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/2893 against CD52
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/732 Antibody-dependent cellular cytotoxicity [ADCC]
• CPC Class Codes: C07K 2317/734 Complement-dependent cytotoxicity [CDC]
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

The present invention relates to optimized CD20 antibodies having Fc variants, methods for their generation, and method for their application, such as methods of enhancing macrophage activation, particularly for therapeutic purposes.

Claims

1. A method of enhancing macrophage activation comprising administering to a patient an anti-CD20 antibody comprising an Fc variant of the human Fc polypeptide comprising SEQ ID NO:

1, wherein said Fc variant portion comprises at least one substitution at a position corresponding to the position of SEQ ID NO;

1 selected from the group consisting of positions 234, 235, 239, 240, 241, 243, 244, 245, 247, 262, 263, 264, 265, 266, 267, 269, 296, 297, 298, 299, 313, 325, 326, 327, 328, 329, 330, 332, 333, and 334, wherein said Fc variant enhances macrophage activation as compared to the unsubstituted Fc variant.

2. The method of enhancing macrophage activation according to claim 1, wherein said Fc variant has the formula:

Vb(234)-Vb(235)-Fx(236-238)-Vb(239)-Vb(240)-Vb(241)-Fx(242)-Vb(243)-Vb(244)-Vb(245)-Fx(246)-Vb(247)-Fx(248-261)-Vb(262)-Vb(263)-Vb(264)-Vb(265)-Vb(266)-Vb(267)-Fx(268)-Fx(269-295)-Vb(296)-Vb(297)-Vb(298)-Vb(299)-Fx(300-312)-Vb(313)-Fx(314-324)-Vb(325)-Fx(326)-Vb(327)-Vb(328)-Vb(329)-Vb(330)-Fx(331)-Vb(332);

wherein;

Vb(234) is selected from the group consisting of L, D, E, N, Q, T, H, Y, I, V, F;

Vb(235) is selected from the group consisting of L, D, S, N, Q, T, H, Y, I, V, F;

Fx(236-238) is (G-G-P);

Vb(239) is selected from the group consisting of S, D, E, N, Q, F, T, H, Y;

Vb(240) is selected from the group consisting of V, I, A, T, M;

Vb(241) is selected from the group consisting of F, W, L, Y, E, R;

Fx(242) is L;

Vb(243) is selected from the group consisting of F, W, L Y, R, Q;

Vb(244) is selected from the group consisting of P, H;

Vb(245) is selected from the group consisting of P, A;

Fx(246) is K;

Vb(247) is selected from the group consisting of P, G, V;

Fx(248-261) is the sequence -(K-D-T-L-M-I-S-R-T-P-E-V-T-C)-;

Vb(262) is selected from the group consisting of V, I, A, T, E;

Vb(263) is selected from the group consisting of V, A, I, M, T;

Vb(264) is selected from the group consisting of V, L, I, W, T, R, F, M, Y, E;

Vb(265) is selected from the group consisting of D, N, Q, Y, F, V, I, L, H, T;

Vb(266) is selected from the group consisting of V, A, I, M, T;

Vb(267) is selected from the group consisting of S, Q, L;

Fx (268) is H;

Vb(269) is selected from the group consisting of E, H, Y, F, R;

Fx (270-295) is the sequence (-D-P-E-V-K-F-N-W-Y-V-D-G-V-E-V-H-N-A-K-T-K-P-R-E-E-Q-);

Vb(296) is selected from the group consisting of Y, E, Q, D, N, S, T, L, I, H;

Vb(297) is selected from the group consisting of N, S, D, E;

Vb(298) is selected from the group consisting of S and H;

Vb(299) is selected from the group consisting of T, I, L, A, S, V, H, F, E;

Fx(300-312) is the sequence (-Y-R-V-V-S-V-L-T-V-L-H-Q-D-);

Vb(313) is selected from the group consisting of W, F;

Fx(314-324) is (L-N-G-K-E-Y-K-C-K-V-S-);

Vb(325) is selected from the group consisting of N, Q, L, I, D, E, A, T, V, H;

Fx(326) is K;

Vb(327) is selected from the group consisting of A, N, L;

Vb(328) is selected from the group consisting of L, M, D, E, N, Q, F, I, V, T, H, A;

Vb(329) is selected from the group consisting of P, F;

Vb(330) is selected from the group consisting of A, L, Y, V, I, F, R, H; and

Fx(331) is P; and

Vb(332) is selected from the group consisting of 1, D, E, N, Q, T, H, Y, and A.

3. The method according to claim 1, wherein said Fc variant has enhanced FcgRIIIa and FcgRI binding.

4. The method according to claim 1, wherein administering said antibody causes increased macrophage phagocytosis.

5. A method of treating a tumor comprising administering to a patient the anti-CD20 antibody of of claim 1.

6. The method of claim 1, wherein said Fc variant comprises at least one substitution at a position corresponding to a position of the human sequence selected from the group consisting of:

240, 244, 245, 247, 262, 263, 266, 299, 313, 325, 328, and 332.

7. The method of claim 6, wherein said Fc variant comprises a substitution at position 332.

8. The method according to claim 1, wherein said Fc variant is selected from the group consisting of 264L, 264I, 241W, 241L, 243W, 243L, 241L/243L/262I/264I, 241W/243W, 241W/243W/262A/264A, 241L/262I, 243L/264I, 243L/262I/264W, 241Y/243Y/262T/264T, 241E/243R/262E/264R, 241E/243Q/262T/264E, 241R/243Q/262T/264R, 241E/243Y/262T/264R, 328M, 328E, 328F, 332E, 328M/332E, 244H, 245A, 247V, 313F, 244H/245A/247V, 247G, 264I/332E, 241E/243R/262E/264R/332E, 241E/243Q/262T/264E/332E, 241R/243Q/262T/264R/332E, 241E/243Y/262T/264R/332E, 298A/332E, 239E/332E, 239Q/332E, 239E, 265G, 265N, 239E/265G, 239E/265N, 239E/265Q, 296E, 296Q, 299I, 327N, 267Q/327S, 267L/327S, 327L, 329F, 330L, 330Y, 332D, 297S, 297D, 297S/332E, 297D/332E, 297E/332E, 265Y/297D/332E, 265Y/297D/299L/332E, 265F/297E/332E, 328I/332E, 328Q/332E, 332N, 332Q, 264T, 264F, 240I, 263I, 266I, 299A, 299S, 299V, 325Q, 325L, 325I, 239D, 239N, 239F, 239D/332D, 239D/332E, 239D/332N, 239D/332Q, 239E/332D, 239E/332N, 239E/332Q, 239N/332D, 239N/332E, 239N/332N, 239N/332Q, 239Q/332D, 239Q/332N, 239Q/332Q, 296D, 296N, 241Y/243Y/262T/264T/297D/332E, 330Y/332E, 264I/330Y/332E, 330L/332E, 264I/330L/332E, 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 235D, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235F, 239T, 239H, 239Y, 240A, 240T, 240M, 263A, 263T, 263M, 264M, 264Y, 266A, 266T, 266M, 269H, 269Y, 269F, 269R, 296S, 296T, 296L, 296I, 298H, 299H, 330V, 330I, 330F, 330R, 330H, 325D, 325E, 325A, 325T, 325V, 325H, 328D/332E, 328E/332E, 328N/332E, 328Q/332E, 328V/332E, 328T/332E, 328H/332E, 328I/332E, 328A, 332T, 332H, 332Y, 332A, 239E/264I/332E, 239Q/264I/332E, 239E/264I/330Y/332E, 239E/264I/298A/330Y/332E, 239D/297D/332E, 239E/297D/332E, 239D/265V/297D/332E, 239D/265I/297D/332E, 239D/265L/297D/332E, 239D/265F/297D/332E, 239D/265Y/297D/332E, 239D/265H/297D/332E, 239D/265T/297D/332E, 264E/297D/332E, 296D/297D/332E, 296E/297D/332E, 296N/297D/332E, 296Q/297D/332E, 296H/297D/332E, 296T/297D/332E, 297D/299V/332E, 297D/299I/332E, 297D/299L/332E, 297D/299F/332E, 297D/299H/332E, 297D/299E/332E, 297D/330Y/332E, 297D/298A/330Y/332E, 239D/330Y/332E, 239N/330Y/332E, 239D/330L/332E, 239N/330L/332E, 264I/298A/332E, 239D/298A/332E, 239N/298A/332E, 239D/264I/332E, 239D/264I/298A/332E, and 239D/264I/330L/332E.

9. The method according to claim 3, wherein said FcgRIIIa is a V158 or F158 allotype of FcgRIIIa.