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JP-7857236-B2 - Immune-activating Fc domain binding molecule

JP7857236B2JP 7857236 B2JP7857236 B2JP 7857236B2JP-7857236-B2

Inventors

  • アマン， マリア
  • ホーファー， トーマス
  • クライン， クリスティアン
  • ラウナー， ラウラ
  • ルクレール， ステファン
  • メスナー， エッケハルト
  • ノイマン， クリスティアーヌ
  • ウマーニャ， パブロ
  • スロウカ， マルレナ
  • ブランシ， アリ
  • カーピー グティエレス チルロス， アレハンドロ
  • クラウス， クリスティーナ
  • コダッリ ディーク， ラウラ
  • ダロフスキ， ダイアナ
  • ファウチ， タニヤ
  • フェッラーラ コラー， クラウディア
  • フライモサー－グルントショーバー， アン
  • ヘルター， シルビア

Assignees

  • エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト

Dates

Publication Date
20260512
Application Date
20210617
Priority Date
20200619

Claims (20)

  1. An immunoactivating fragment crystallizable (Fc) domain binding molecule, (a) an Fc domain binding moiety that specifically binds to a target Fc domain containing at least one amino acid substitution of the first set, (b) Immune activation portion and (c) Half-life extension Fc domain and Includes, The Fc domain binding portion is specifically capable of binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), The half-life extension Fc domain comprises at least one amino acid substitution of the second set, At least one amino acid substitution in the second set includes a substitution at position P329 (numbered according to Kabat's EU index) by an amino acid selected from the list consisting of arginine (R), leucine (L), isoleucine (I), and alanine (A), An immune-activating Fc domain binding molecule.
  2. The immunoactivating Fc domain binding molecule according to claim 1, wherein the Fc domain binding portion does not specifically bind to the half-life extension Fc domain.
  3. The immunoactivating Fc domain binding molecule according to claim 1 or 2, wherein at least one amino acid substitution in the first set reduces the binding affinity and/or effector function to the Fc receptor, and at least one amino acid substitution in the second set includes one or more amino acid substitutions at the same amino acid position as at least one amino acid substitution in the first set, and the amino acids of at least one amino acid substitution in the second set are substituted with different amino acids at the same position as at least one amino acid substitution in the first set.
  4. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 3 , wherein at least one amino acid substitution of the first set comprises at least one amino acid substitution at a position selected from the list consisting of 233, 234, 235, 238, 253, 265, 269, 270, 297, 310, 331, 327 and 435 (numbering according to Kabat's EU index).
  5. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 4 , wherein at least one amino acid substitution of the second set comprises at least one amino acid substitution at a position selected from the list consisting of 233, 234, 235, 238, 253, 265, 269, 270, 297, 310, 331, 327 and 435 (numbering according to Kabat's EU index).
  6. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 5 , wherein at least one amino acid substitution of the second set comprises a substitution by arginine (R) at position P329 (numbered according to Kabat's EU index).
  7. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 6, wherein the Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing an amino acid substitution P329G (numbering according to Kabat's EU index), but is not capable of specific binding to a parental non-mutant IgG1 Fc domain.
  8. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 7 , wherein at least one amino acid substitution of the first set comprises at least one amino acid substitution at a position selected from the group L234, L235 (Kabat's EU index numbering).
  9. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 8 , wherein at least one amino acid substitution of the second set comprises at least one amino acid substitution at a position selected from the group L234, L235 (Kabat's EU index numbering).
  10. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 6, wherein the target Fc domain comprises three amino acid substitutions that reduce binding to and/ or effector function of the activated Fc receptor, the amino acid substitutions being L234A, L235A, and P329G (Kabat's EU index numbering).
  11. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 10, wherein the half-life extension Fc domain comprises three amino acid substitutions that reduce binding to and/or effector function of the activated Fc receptor, the amino acid substitutions being L234A, L235A, and P329X (Kabat's EU index numbering), where X is an amino acid other than glycine (G).
  12. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 11 , wherein the Fc domain binding portion and/or the immunoactivating portion is a Fab molecule.
  13. The Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion, (i) Heavy chain variable region (VH), (a) Heavy chain complementarity determining region (CDR H) 1 amino acid sequence RYWMN (SEQ ID NO: 1); (b) CDR H2 amino acid sequence selected from the group consisting of EITPDSSTINYTPSLKD (SEQ ID NO: 2), EITPDSSTINYTPSLKG (SEQ ID NO: 11), and EITPDSSTINYAPSLKG (SEQ ID NO: 16); and (c) CDR H3 amino acid sequence PYDYGAWFAS (SEQ ID NO: 3) The heavy chain variable region (VH), (ii) Light chain variable region (VL), (d) Light chain complementarity determining region (CDR L) 1 amino acid sequence RSSTGAVTTTSNYAN (SEQ ID NO: 4); (e) CDR L2 amino acid sequence GTNKRAP (SEQ ID NO: 5); and (f) CDR L3 amino acid sequence ALWYSNHWV (SEQ ID NO: 6) An immunoactivating Fc domain binding molecule according to any one of claims 1 to 12 , comprising a light chain variable region (VL).
  14. The Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion, (i) Heavy chain variable region (VH), (a) Heavy chain complementarity determining region (CDR H) 1 amino acid sequence RYWMN (SEQ ID NO: 1); (b) CDR H2 amino acid sequence EITPDSSTINYTPSLKD (SEQ ID NO: 2); and (c) CDR H3 amino acid sequence PYDYGAWFAS (SEQ ID NO: 3) The heavy chain variable region (VH), (ii) Light chain variable region (VL), (d) Light chain complementarity determining region (CDR L) 1 amino acid sequence RSSTGAVTTTSNYAN (SEQ ID NO: 4); (e) CDR L2 amino acid sequence GTNKRAP (SEQ ID NO: 5); and (f) CDR L3 amino acid sequence ALWYSNHWV (SEQ ID NO: 6) The immunoactivating Fc domain binding molecule according to claim 13 , comprising a light chain variable region (VL).
  15. The Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion, (i) Heavy chain variable region (VH), (a) Heavy chain complementarity determining region (CDR H) 1 amino acid sequence RYWMN (SEQ ID NO: 1); (b) CDR H2 amino acid sequence EITPDSSTINYTPSLKG (SEQ ID NO: 11); and (c) CDR H3 amino acid sequence PYDYGAWFAS (SEQ ID NO: 3) The heavy chain variable region (VH), (ii) Light chain variable region (VL), (d) Light chain complementarity determining region (CDR L) 1 amino acid sequence RSSTGAVTTTSNYAN (SEQ ID NO: 4); (e) CDR L2 amino acid sequence GTNKRAP (SEQ ID NO: 5); and (f) CDR L3 amino acid sequence ALWYSNHWV (SEQ ID NO: 6) The immunoactivating Fc domain binding molecule according to claim 13 , comprising a light chain variable region (VL).
  16. The Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion, (i) Heavy chain variable region (VH), (a) Heavy chain complementarity determining region (CDR H) 1 amino acid sequence RYWMN (SEQ ID NO: 1); (b) CDR H2 amino acid sequence EITPDSSTINYAPSLKG (SEQ ID NO: 16); and (c) CDR H3 amino acid sequence PYDYGAWFAS (SEQ ID NO: 3) The heavy chain variable region (VH), (ii) Light chain variable region (VL), (d) Light chain complementarity determining region (CDR L) 1 amino acid sequence RSSTGAVTTTSNYAN (SEQ ID NO: 4); (e) CDR L2 amino acid sequence GTNKRAP (SEQ ID NO: 5); and (f) CDR L3 amino acid sequence ALWYSNHWV (SEQ ID NO: 6) The immunoactivating Fc domain binding molecule according to claim 13 , comprising a light chain variable region (VL).
  17. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 16, wherein the Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing an amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion comprises a heavy chain variable region sequence which is at least 95 %, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs. 7, SEQ ID NOs. 12, SEQ ID NOs. 17, and SEQ ID NOs. 19, and a light chain variable region sequence which is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs. 8 and SEQ ID NOs. 13.
  18. The Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing the amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion, (i) A heavy chain variable region sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence number 7, and a light chain variable region sequence that is at least 95 %, 96%, 97%, 98%, 99%, or 100% identical to sequence number 8, (ii) A heavy chain variable region sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to sequence number 12, and a light chain variable region sequence that is at least 95 %, 96%, 97%, 98%, 99%, or 100% identical to sequence number 13, (iii) A heavy chain variable region sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, and a light chain variable region sequence that is at least 95 %, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 13, or (iv) A heavy chain variable region sequence that is at least 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 19, and a light chain variable region sequence that is at least 95 %, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 13. The immunoactivating Fc domain binding molecule according to claim 17 .
  19. The immunoactivating Fc domain binding molecule according to claim 18, wherein the Fc domain binding portion is capable of specific binding to an IgG1 Fc domain containing an amino acid substitution P329G (numbering according to Kabat's EU index), and the Fc domain binding portion includes the heavy chain variable region sequence of SEQ ID NO: 19 and the light chain variable region of SEQ ID NO: 13 .
  20. The immunoactivating Fc domain binding molecule according to any one of claims 1 to 19 , wherein the Fc domain binding portion comprises a first Fab molecule and the immunoactivating portion comprises a second Fab molecule.

Description

Field of Invention The present invention generally relates to novel immunoactivating Fc domain binding molecules for the activation of immune cells and redirection to specific target cells. Furthermore, the present invention relates to polynucleotides encoding such molecules, as well as vectors and host cells containing such polynucleotides. The present invention further relates to methods for producing the bispecific antigen-binding molecules of the present invention, and methods for using these bispecific antigen-binding molecules in the treatment of diseases. Background: Selective destruction of individual cells or specific cell types is often desirable in a variety of clinical situations. For example, the primary goal of cancer treatment is to specifically destroy tumor cells while leaving healthy cells and tissues intact and undamaged, or to destroy specific cell subsets identified by specific surface antigens. An attractive way to achieve this is to induce an immune response against target cells by recruiting immune effector cells such as natural killer (NK) cells, monocytes/macrophages, or cytotoxic T lymphocytes (CTLs) to attack and destroy tumor cells. One method for inducing immune effector cell-mediated target cell death or depletion is antibody-dependent cytotoxicity (ADCC) or antibody-dependent cytotoxicity (ADCP) mediated by ADCC-competent antibodies of IgG1 isotypes and antibodies with enhanced ADCC effector function (Zahavi et al, Antibody Therapeutics, 1, 7-12 (2018)). Alternatively, T cells may be recruited to kill target cells via (T cell) bispecific antibodies designed to bind to surface antigens on target cells and have a second binding site to the invariant component of the T cell receptor (TCR) complex that activates it (Clynes and Desjarlais, Annú Rev Med 70:427–450 (2019)). Several bispecificity formats have been developed, including BiTE (bispecific T cell engagers) (Nagorsen and Baeuerle, Exp Cell Res 317, 1255-1260 (2011)), diabodynes (Holliger et al., Prot Eng 9, 299-305 (1996)), DART (biaffinity retargeting) (Moore et al., Blood 117, 4542-51 (2011)), or so-called 2+1 T cell bispecific antibodies (TCBs) (Bacac et al., Clin Cancer Res 24, 4785-4797 (2018)), and their suitability for T cell-mediated immunotherapy is being investigated. The various formats under development demonstrate significant potential stemming from the redirection and activation of immune cells in immunotherapy. To date, developed bispecific antibodies consistently engage directly with the desired antigen, thereby linking target cells to CTLs and resulting in target cell lysis. These bispecific antibody formats face challenges related to toxicity, applicability, and productivity. Furthermore, for each single target (combination), it is necessary to construct individual molecules specific to each target. The therapeutic utility of antibodies and their derivatives is not limited to their function as T-cell engagers; indications are also found in the modulation of checkpoints of inhibitors or activators. Exemplarily, the use of immune checkpoint inhibitor antibodies has demonstrated persistent responses in several indications (Hodie et al. N Engl J Med.; 363(8):711-23. (2010); Prieto PA, et al. Clin cancer Res.; 18:2039-2047 (2012)). More recently, it has been shown that the activity of T cell bispecific antibodies can be further enhanced by bispecific substances that activate so-called costimulatory pathways on T cells via the activation of CD28 (Skokos et al., Sci Trans Med 12(525):1-14(2020)) or 4-1BB signaling (Claus et al., Sci Trans Med 11(496), eaav5989(2019)). In addition to current successes, therapies are limited in their flexibility to target multiple different antigens and their ability to selectively utilize combined NK or CTL functions for a single application. Summary of the Invention: An immunoactivating fragment crystallizable (Fc) domain binding molecule, (a) an Fc domain binding moiety that specifically binds to a target Fc domain containing at least one amino acid substitution of the first set, (b) An immune-activating Fc domain binding molecule comprising an immune-activating moiety is provided herein. In one embodiment, at least one amino acid substitution in the first set reduces binding to the Fc receptor and/or diminishes effector function. In one embodiment, the immune-activating Fc domain binding molecule is (c) further comprising a half-life extension Fc domain, The Fc domain binding portion does not specifically bind to the half-life extension Fc domain. In one embodiment, the half-life extension Fc domain contains at least one amino acid of a second set. In one embodiment, the substitution of at least one amino acid of the second set reduces binding to Fc. In one embodiment, the target Fc domain and/or the half-life extension Fc domain is composed of first and second subunits capable of stable association. In one embodiment, the target Fc domain and/or half-life extension Fc domain