Selective heterodimeric Fc variant for Fc gamma RIIB

JP7875129B2Active Publication Date: 2026-06-17ZYMEWORKS BC INC
View PDF 11 Cites 0 Cited by

Patent Information

Authority / Receiving Office
JP · JP
Patent Type
Patents
Current Assignee / Owner
ZYMEWORKS BC INC
Filing Date
2021-05-20
Publication Date
2026-06-17

Smart Images

  • Figure 0007875129000348
    Figure 0007875129000348
  • Figure 0007875129000349
    Figure 0007875129000349
  • Figure 0007875129000350
    Figure 0007875129000350
Patent Text Reader

Abstract

Provided are heterodimeric Fc variants, polypeptides comprising the heterodimeric Fc variants, and polynucleotides encoding the heterodimeric Fc variants, which comprise one or more asymmetric amino acid mutations in the CH2 domain and have increased binding selectivity to FcγRIIb compared to the parent Fc region. The one or more asymmetric mutations comprise a substitution of a loop in the CH2 domain, a mutation at position 236 in the CH2 domain, or a combination of a substitution of a loop in the CH2 domain and a mutation at position 236 in the CH2 domain.
Need to check novelty before this filing date? Find Prior Art

Claims

1. A heterodimer Fc variant comprising a first Fc polypeptide and a second Fc polypeptide, One of the Fc polypeptides includes substitution of amino acids 325 to 331 by a polypeptide having a length of 8 to 15 amino acids. The aforementioned heterodimer Fc variant exhibits increased selectivity for binding to FcγRIIb compared to the parent Fc region. The heterodimer Fc variant is a variant of immunoglobulin G1 (IgG1) Fc, and the parent Fc region is wild-type IgG1 Fc. The polypeptides mentioned above are sequence numbers 6, 8, 9, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, and 52. , containing an amino acid sequence described in any one of 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90, The amino acid numbering follows the EU index. The aforementioned heterodimer Fc variant.

2. The heterodimer Fc variant according to claim 1, wherein the polypeptide comprises an amino acid sequence described in any one of SEQ ID NOs: 6, 8, 47, 68, or 73.

3. The heterodimer Fc variant according to claim 1 or 2, further comprising a symmetric mutation at position 236 of the first and second Fc polypeptides, wherein the mutation at position 236 is selected from G236D, G236N, and G236K.

4. The heterodimer Fc variant according to claim 3, wherein the mutation at position 236 is G236D or G236N.

5. The above substitutions of amino acids 325-331 are present in the second Fc polypeptide. The first Fc polypeptide includes a mutation at position 236 selected from G236A, G236D, G236E, G236F, G236H, G236I, G236L, G236N, G236P, G236Q, G236S, G236T, G236V, G236W and G236Y, and the second Fc polypeptide includes the mutation G236D. The heterodimer Fc variant according to claim 1 or 2.

6. The substitutions of amino acids 325 to 331 are present in the second Fc polypeptide, The first Fc polypeptide comprises G236N, and the second Fc polypeptide comprises a mutation at position 236 selected from G236D, G236E, G236K, G236N, and G236T. The heterodimer Fc variant according to claim 1 or 2.

7. The substitutions of amino acids 325 to 331 are present in the second Fc polypeptide, The first Fc polypeptide contains mutant G236N, and the second Fc polypeptide contains mutant G236D. The heterodimer Fc variant according to claim 1 or 2.

8. The heterodimer Fc variant according to any one of claims 1 to 7, wherein the substitution of amino acids 325 to 331 is in the second Fc polypeptide, and the second Fc polypeptide further comprises one or more mutations selected from S239D, S239E, V266I, V266L, S267A, S267I, S267V, S267Q and H268D.

9. The heterodimer Fc variant according to any one of claims 1 to 7, wherein the substitution of amino acids 325 to 331 is in the second Fc polypeptide, and the second Fc polypeptide further comprises (i) mutation S239D or S239E, and / or (ii) mutation H268D, and / or (iii) mutation S267A, S267I, or S267V.

10. The heterodimer Fc variant according to claim 9, wherein the second Fc polypeptide comprises mutants S239D, H268D, and S267V.

11. The above substitutions of amino acids 325-331 are present in the second Fc polypeptide. The first Fc polypeptide further comprises a mutation at one or more of the positions 234, 235, 237, and 239, (i) The variation at position 234 is selected from L234A, L234D, L234E, L234F, L234G, L234H, L234I, L234N, L234P, L234Q, L234S, L234T, L234V, L234W and L234Y, (ii) The variation at position 235 is selected from L235A, L235D, L235E, L235F, L235H, L235I, L235N, L235P, L235Q, L235S, L235T, L235V, L235W and L235Y, (iii) The variation at position 237 is selected from G237A, G237D, G237F, G237H, G237L, G237N, G237P, G237S, G237V, G237W and G237Y, (iv) The variation at position 239 is selected from S239A, S239D, S239E, S239F, S239G, S239H, S239I, S239L, S239N, S239Q, S239R, S239T, S239V, S239W and S239Y. The heterodimer Fc variant according to any one of claims 1 to 10.

12. The substitutions of amino acids 325 to 331 are present in the second Fc polypeptide, The first Fc polypeptide further comprises (i) mutations L235D, L235E, L235F, or L235T, and / or (ii) mutations G237A or G237D. The heterodimer Fc variant according to any one of claims 1 to 10.

13. The above substitutions of amino acids 325-331 are present in the second Fc polypeptide. The second Fc polypeptide further comprises a mutation at one or more of the positions 234, 235, 237, 240, 263, 264, 266, 269, 271, 273, 323, and 332, (i) The variation at position 234 is selected from L234A, L234E, L234F, L234G, L234H, L234I, L234K, L234N, L234P, L234Q, L234S, L234T, L234V, L234W and L234Y, (ii) The variation at position 235 is selected from L235A, L235D, L235F, L235G, L235N, L235S, L235W and L235Y, (iii) The variation at position 237 is selected from G237F, G237I, G237K, G237L, G237Q, G237T, G237V and G237Y, (iv) The mutation at position 240 is selected from V240I and V240L, (v) The variation at position 263 is V263T, (vi) The variation at position 264 is V264T, (vii) The mutation at position 266 is V266I, (viiii) The mutation at position 269 is E269Q, The mutation at position (ix) 271 is P271D, (x) The variation at position 273 is selected from V273A and V273I, (xi) The variation at position 323 is selected from V323A and V323I, (xii) The variation at position 332 is selected from I332F and I332L. A heterodimer Fc variant according to any one of claims 1 to 12.

14. The substitutions of amino acids 325 to 331 are present in the second Fc polypeptide, The second Fc polypeptide further comprises one or more of the mutations G237F, S239D, S267V, H268D, and I332L. A heterodimer Fc variant according to any one of claims 1 to 12.

15. The substitutions of amino acids 325 to 331 are present in the second Fc polypeptide, The second Fc polypeptide further comprises the mutations G237F, S239D, S267V, and H268D. A heterodimer Fc variant according to any one of claims 1 to 12.

16. The heterodimer Fc variant according to claim 15, wherein the second Fc polypeptide further comprises mutant I332L.

17. The heterodimer Fc variant according to claim 1, wherein the heterodimer Fc variant includes an amino acid mutation described in any one of the variants shown in Table 6.22, Table 6.24, Table 6.25, or Table 6.

27.

18. (i) The first Fc polypeptide contains mutation G236N_G237D, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D (Variant 31186), (ii) The first Fc polypeptide contains mutation L235F_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D (Variant 31187), (iii) The first Fc polypeptide contains mutation L235F_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (G330 ＊ K) + G236D_G237F_S239D_S267V_H268D (Variant 31188) or (iv) The first Fc polypeptide contains mutation G236N_G237D, and the second Fc polypeptide contains mutation template 7 (E328 ＊ H_E329 ＊ R_A331 ＊ BY) + G236D_G237F_S239D_S267V_H268D (Variant 31191) or (v) The first Fc polypeptide contains mutation L235F_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 31213), (vi) The first Fc polypeptide contains mutation L235F_G236N_G237A_T250V_A287F, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_T250V_S267V_H268D_A287F (Variant 31274), (vii) The first Fc polypeptide comprises the mutation L235F_G236N_G237A_T250V_M428F, and the second Fc polypeptide comprises the mutation template 1 (D329 ＊ I)+G236D_G237F_S239D_T250V_S267V_H268D_M428F (variant 31275), or (viiii) The first Fc polypeptide contains mutation L235F_G236N_G237A_A287F_M428F, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_A287F_M428F (variant 31276), (ix) The first Fc polypeptide contains mutation G236N_G237D, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32210), (x) The first Fc polypeptide contains mutation G236N_G237E, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32211), (xi) The first Fc polypeptide contains mutation G236N, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32212), (xi) The first Fc polypeptide contains mutation L235D_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (variant 32226), (xiii) The first Fc polypeptide contains mutation L235E_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (variant 32227), (xiv) The first Fc polypeptide contains mutation L235V_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32230), (xv) The first Fc polypeptide contains mutation L235Y_G236N_G237A, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32231), (xvi) The first Fc polypeptide contains the mutation G236N_G237A_S239P, and the second Fc polypeptide contains the mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (variant 32242), (xvii) The first Fc polypeptide contains mutation L234D_G236N_G237A, and the second Fc polypeptide contains mutation template 7+G236D_G237F_S239D_S267V_H268D (variant 32282), (xviiii) The first Fc polypeptide contains mutation L235D_G236N_G237A, and the second Fc polypeptide contains mutation template 7+G236D_G237F_S239D_S267V_H268D (variant 32284), (xix) The first Fc polypeptide contains the mutation G236N_G237A_S239G, and the second Fc polypeptide contains the mutation template 7+G236D_G237F_S239D_S267V_H268D (variant 32287), (xx) The first Fc polypeptide contains the mutation G236N_G237A_S239H, and the second Fc polypeptide contains the mutation template 7+G236D_G237F_S239D_S267V_H268D (variant 32288), (xxi) The first Fc polypeptide contains the mutation G236N_G237E, and the second Fc polypeptide contains the mutation template 7+G236D_G237F_S239D_S267V_H268D (variant 32296), (xxii) The first Fc polypeptide contains mutation L234F_L235D_G236N_H268Q_A327G_A330K_P331S, and the second Fc polypeptide contains mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D (Variant 31192), (xxiii) The first Fc polypeptide contains the mutation L234F_L235D_G236N_H268Q_A327G_A330K_P331S, and the second Fc polypeptide contains the mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32292), (xxiv) The first Fc polypeptide contains the mutation L234F_G236N_S267A_H268Q_A327G_A330K_P331S, and the second Fc polypeptide contains the mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32293), (xxv) The first Fc polypeptide contains the mutation L234F_G236N_H268Q_A327G_A330T_P331S, and the second Fc polypeptide contains the mutation template 1 (D329 ＊ I) Includes +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32294), or (xxvi) The first Fc polypeptide contains the mutation L234F_G236N_H268Q_A327G_P329I_A330K_P331S, and the second Fc polypeptide contains the mutation template 1 (D329 ＊ I) Including +G236D_G237F_S239D_S267V_H268D_I332L (Variant 32295), The heterodimer Fc variant according to claim 1.

19. The heterodimer Fc variant according to any one of claims 1 to 18, wherein the first Fc polypeptide and the second Fc polypeptide further comprise one or more mutations selected from A287F, T250V, L309Q, and M428F.

20. The heterodimer Fc variant according to claim 19, wherein the first Fc polypeptide and the second Fc polypeptide further comprise the mutant A287F / M428F, A287F / T250V, M428F / T250V, or T250V / L309Q.

21. A heterodimer Fc variant according to any one of claims 1 to 20, which is a variant of human IgG1 Fc.

22. The selectivity of the heterodimer Fc variant to binding to FcγRIIb is increased by at least 1.5 times compared to the parent Fc region, where, FcγRIIb selectivity ratio (Fold Difference) = FcγRIIb affinity ratio / FcγRIIaR affinity ratio ratio And, During the ceremony, FcγRIIb affinity factor difference = K Ｄ FcγRIIb (parent) / K Ｄ FcγRIIb (variant) and FcγRIIaR affinity factor difference = K Ｄ FcγRIIaR (parent) / K Ｄ FcγRIIaR (variant) The heterodimer Fc variant according to any one of claims 1 to 21.

23. A heterodimer Fc variant according to any one of claims 1 to 22, wherein the binding affinity to FcγRIIb is increased compared to the parent Fc region.

24. The binding affinity of the heterodimer Fc variant to FcγRIIb is increased by at least 10 times compared to the parent Fc region, where, FcγRIIb affinity factor difference = K Ｄ FcγRIIb (parent) / K Ｄ FcγRIIb (variant) The heterodimer Fc variant according to claim 23.

25. A polypeptide comprising a heterodimer Fc variant according to any one of claims 1 to 24, and one or more antigen-binding domains fused to or covalently bonded to the heterodimer Fc variant.

26. The polypeptide according to claim 25, wherein at least one of the antigen-binding domains binds to a tumor-associated antigen or a tumor-specific antigen.

27. A pharmaceutical composition comprising a heterodimer Fc variant according to any one of claims 1 to 24 or a polypeptide according to claim 25 or 26, and a pharmaceutically acceptable carrier or diluent.

28. Use of the polypeptide according to claim 26 in the manufacture of a drug for the treatment of cancer.

29. A nucleic acid encoding a heterodimer Fc variant according to any one of claims 1 to 24, or a polypeptide according to claim 25 or 26.

30. A host cell comprising the nucleic acid described in claim 29.

31. A method for preparing a heterodimer Fc variant according to any one of claims 1 to 24, or a polypeptide according to claim 25 or 26, the method comprising expressing a nucleic acid encoding the heterodimer Fc variant or the polypeptide in a host cell.