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JP-7856312-B2 - Conditionally active anti-HER2 antibodies, antibody fragments, their immunoconjugates, and their use

JP7856312B2JP 7856312 B2JP7856312 B2JP 7856312B2JP-7856312-B2

Inventors

  • ショート ジェイ エム
  • フレイ ガーハード
  • チャング フウェイ ウェン

Assignees

  • バイオアトラ インコーポレイテッド

Dates

Publication Date
20260511
Application Date
20210122
Priority Date
20200123

Claims (20)

  1. An antibody that binds to the HER2 protein or an antigen-binding antibody fragment thereof, wherein the binding affinity to the HER2 protein at pH 5.0 to 6.8 is higher than the binding affinity to the HER2 protein at pH 7.0 to 7.6. The antibody or its antigen-binding antibody fragment comprises a heavy chain variable region and a light chain variable region, The heavy chain variable region includes three complementarity-determining regions having sequences H1, H2, and H3. The H1 sequence is GFX 1 IKDTYIH (Sequence ID 1), The H2 sequence is X 2 IX 3 PTX 4 X 5 YX 6 X 7 YADSVKG (Sequence ID 2), The H3 sequence is WGGDGFYX 8 MDY (Sequence ID 3), In the formula, X1 is N or W, X2 is R or K, X3 is Y, K or D, X4 is N or A, X5 is G or K , X6 is T or D, X7 is R or E, and X8 is A or E. The light chain variable region includes three complementarity-determining regions having sequences L1, L2, and L3. The L1 sequence is RASQDVNTX 9VA (sequence number 4), The L2 sequence is SASFLYS (Sequence ID 5), The L3 sequence is QQX 10 YTTPPPT (sequence number 6), In the formula, X9 is A or D, and X10 is H, D or E. However, if X1 to X8 are N, R, Y, N, G, T, R, and A respectively, then X9 is not A, and X10 is not H. If X1 is W, then X2 to X10 are not R, Y, N, G, T, R, A, A, and H respectively. If X3 is K or D, then X1 to X2 and X4 to X10 are not N, R, N, G, T, R, A, A, and H, respectively. If X4 is A, then X1 to X3 and X5 to X10 are not N, R, Y, G, T, R, A, A, and H, respectively. If X7 is E, then X1 to X6 and X8 to X10 are not N, R, Y, N, G, T, A, A, and H respectively, and if X8 is E, then X1 to X6 and X8 to X10 are not N, R, Y, N, G, T, R, A, and H respectively, The antibody or antigen-binding antibody fragment, compared to a parent antibody or antibody fragment where X1 to X10 are N, R, Y, N, G, T, R, A, A, and H respectively, has a single substitution in one of the H1, H2, and H3 sequences in the heavy chain variable region, and up to two substitutions in the L1 and L3 sequences in the light chain variable region. An antibody or an antigen-binding antibody fragment thereof.
  2. A bispecific antibody or antigen-binding antibody fragment comprising the antibody or antigen-binding antibody fragment described in claim 1, wherein the bispecific antibody or antigen-binding antibody fragment also binds to the CD3 protein, and the bispecific antibody or antigen-binding antibody fragment further comprises six anti-CD3 complementarity determining regions L4, L5, L6, L7, L8, and L9, The L4 sequence is GFTFNTYAMN (sequence number 54), The L5 sequence is RIRSKYNNYATYYADSVKD (sequence number 55), The L6 sequence is HX 11 NFX 12 NSKVSWFX 13 Y (Sequence ID 70), The L7 sequence is RSSX 14 GAVTTSNYDN (sequence number 71), The L8 sequence is GTNKRAP (sequence number 58), The L9 sequence is ALWYSNLWV (sequence number 59), In the formula, X 11 is G, S, A, or T, X 12 is G or P, X 13 is A or Q, and X 14 is T or A. The binding affinity to CD3 at pH 5.0–6.8 is higher than the binding affinity to CD3 at pH 7.0–7.6. The antibody or its antigen-binding antibody fragment, compared to a parent antibody or its antibody fragment where X1 to X14 are N, R, Y, N, G, T, R, A, A, H, G, G, A, and T respectively, has a single substitution in one of the H1, H2, and H3 sequences in the heavy chain variable region, and up to three substitutions in the L1, L3-L4, and L6-L7 sequences in the light chain variable region. The antibody according to claim 1 or the antigen-binding antibody fragment thereof.
  3. The antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof, according to claim 1 or 2, wherein the H1 sequence is GFWIKDTYIH (SEQ ID NO: 7) or GFNIKDTYIH (SEQ ID NO: 50).
  4. The aforementioned H2 sequence is KIYPTNGYTRYADSVKG (Sequence No. 8) RIKPTNGYTRYADSVKG (Sequence No. 9) RIDPTNGYTRYADSVKG (Sequence No. 10) RIYPTAGYTRYADSVKG (Sequence No. 11) RIYPTNKYTRYADSVKG (Sequence No. 12) RIYPTNGYDRYADSVKG (Sequence No. 13) An antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof, selected from the group consisting of RIYPTNGYTEYADSVKG (SEQ ID NO: 14) and RIYPTNGYTRYADSVKG (SEQ ID NO: 49), according to claim 1 or 2.
  5. The antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof , according to claim 1 or 2, wherein the H3 sequence is WGGDGFYEMDY (SEQ ID NO: 15) or WGGDGFYAMDY (SEQ ID NO: 51).
  6. The antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof , according to claim 1 or 2, wherein the L1 sequence is RASQDVNTDVA (SEQ ID NO: 16) or RASQDVNTAVA (SEQ ID NO: 52).
  7. The antibody or antigen-binding antibody fragment thereof , or a bispecific antibody or antigen-binding antibody fragment thereof, according to claim 1 or 2, wherein the L3 sequence is QQDYTTPPPT (SEQ ID NO: 17), QQEYTTPPPT (SEQ ID NO: 18), or QQHYTTPPPT (SEQ ID NO: 53).
  8. The bispecific antibody or antigen-binding antibody fragment according to claim 2, wherein the L6 sequence is one of sequence numbers 56 and 60-67, and the L7 sequence is sequence number 57, 68, or 69.
  9. An antibody or antigen-binding antibody fragment thereof according to claim 1 or 2, or a bispecific antibody or antigen-binding antibody fragment thereof, wherein the heavy chain variable region is EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAKIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 19) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTEYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 20) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIKPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 21) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIDPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 22) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTAGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 23) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNKYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 24) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYDRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 25) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYEMDYWGQGTLVTVSS (Sequence No. 26) It is one of the following: EVQLVESGGGLLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVS (Sequence No. 27), or EVQLVESGGGLLVQPGGSLRLSCAASGFWIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 28). The light chain variable region is DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 29) DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 30) It is one of the following: DIQMTQSPSSLSASVGDRVTITCCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQDYTTPPTFGQGTKVEIK (Sequence ID 31) or DIQMTQSPSSLSASVGDRVTITCCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQEYTTPPTFGQGTKVEIK (Sequence ID 32). However , if the heavy chain variable region is sequence number 20, 21, 22, 23, 26, 27, or 28, the light chain variable region is not sequence number 29. The aforementioned antibody or its antigen-binding antibody fragment , or a bispecific antibody or its antigen-binding antibody fragment .
  10. The aforementioned heavy chain variable region is EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 33) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAKIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 19) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTEYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 20) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIKPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 21) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIDPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 22) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTAGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 23) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNKYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 24) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYDRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 25) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYEMDYWGQGTLVTVSS (Sequence No. 26) It is one of the following: EVQLVESGGGLLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVS (Sequence No. 27), or EVQLVESGGGLLVQPGGSLRLSCAASGFWIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 28). The aforementioned light chain variable region is DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 30) An antibody or antigen-binding antibody fragment thereof according to claim 1 or 2, or a bispecific antibody or antigen-binding antibody fragment thereof, which is one of DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQDYTTPPTFGQGTKVEIK (SEQ ID NO: 31) or DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQEYTTPPTFGQGTKVEIK (SEQ ID NO: 32).
  11. The heavy chain variable region and the light chain variable region are EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAKIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVSS (Sequence ID 19) and An antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof, selected from DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 29), according to claim 9.
  12. The aforementioned heavy chain variable region is EVQLVESGGGLLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRRWGGDGFYAMDYWGQGTLVTVSS (Sequence ID 33), The aforementioned light chain variable region is DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAAWYQQKPGKAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 30), The antibody or antigen-binding antibody fragment thereof according to claim 10, or a bispecific antibody or antigen-binding antibody fragment thereof, which is one of the following: DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQDYTTPPTFGQGTKVEIK (SEQ ID NO: 31), or DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQEYTTPPTFGQGTKVEIK (SEQ ID NO: 32).
  13. The aforementioned light chain variable region is DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLLIYSASFLYSGVPPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (Sequence ID 30) The aforementioned heavy chain variable region is EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 33), EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVAKIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 19) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTEYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 20) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIKPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 21) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIDPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 22) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTAGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 23) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNKYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 24) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYDRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (Sequence No. 25) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYEMDYWGQGTLVTVSS (Sequence No. 26) EVQLVESGGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVES (Sequence ID 27), or An antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof, which is one of EVQLVESGGGLLVQPGGSLRLSCAASGFWIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAADTSKNTAYLQMNSLRAEDTAVYYCSRWGGGDGFYAMDYWGQGTLVTVSS (Sequence ID 28), according to claim 9.
  14. The aforementioned heavy chain variable region is EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIKPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 35), EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNKYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 36), EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYDRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 37), EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYEMDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPELLGGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 38), or EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYT RYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVT VESASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPA It is one of the sequences PELLGGGPSVFLFPPKPKDTLMISRTPEPEVTCVVVDVSSHEDPEVKFNWYVDGVEVHNAKTKPREEQYQSTYRVVSSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 39), The light chain variable region is DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHSNFGNSKVSWFQYWGQGTLVTVSSGGGSGGGSGGGSGGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 41), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHGNFGNSKVSWFQYWGQGTLVTVSSGGGSGGGSGGGSGGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 42), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHGNFPPNSKVSWFQYWGQGTLVTVSSGGGSGGGSGGGSGGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 43), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHANFGNSKVSWFAYWGQGTLVTVSSGGGSGGGSGGGSGGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 44), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHGNFPPNSKVSWFAYWGQGTLVTVSSGGGSGGGSGGGSGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 45), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLR LSCAASGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHTNFGNSKVSWFAYWGQGTLVTVSSGGGSGGGSGGGSGGQAVVTQEPSLTVSPGGGTVTLTCRSSSTGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 46), DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPP SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSSTLT LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLRL SCAASSGFTFNTYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHSNFGNSKVSWFAYWGQGTLVTVSSGGGSGGGSGGGSGGQAVVTQEPSLTVSPGGGTVTLTCRSSAGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 47), or DIQMTQSPSSLSASVGDRVTITCRASQDVNTDVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRS GTDFTLISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGECSRSGGGGEVQLVESGGGLVQPGGSLRLSCAASGFTFNTYAMNWVRQAPGKGLEWVAR An antibody or antigen-binding antibody fragment thereof , or a bispecific antibody or antigen-binding antibody fragment thereof, which is one of IRSKYNNYATYYADSVKDRFTISRDDSKNSLYLQMNSLKTEDTAVYYCVRHTNFGNSKVSWFAYWGQGTLVVTVSSSGGGSGGGSGGGSGGGSGGQAVVTQEPSLTVSPGGGTVTLTCRSSAGAVTTTSNYDNWVQQKPGQAPRGLIGGTNKRAPWTPARFSGSLLGGKAALTITGAQAEDEADYYCALWYSNLWVFGGGTKLTVLSR (Sequence ID 48), according to claim 1 or 2.
  15. The antibody or antigen-binding antibody fragment thereof, or a bispecific antibody or antigen-binding antibody fragment thereof, according to claim 14, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 35, and the light chain variable region has one of the amino acid sequences of SEQ ID NOs: 41 to 48.
  16. The antibody or its antigen-binding antibody fragment or bispecific antibody or its antigen-binding antibody fragment according to claim 14, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 36, and the light chain variable region has any one of the amino acid sequences of SEQ ID NOs: 41 to 48.
  17. The antibody or its antigen-binding antibody fragment or bispecific antibody or its antigen-binding antibody fragment according to claim 14, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 37, and the light chain variable region has any one of the amino acid sequences of SEQ ID NOs: 41 to 48.
  18. The antibody or its antigen-binding antibody fragment or bispecific antibody or its antigen-binding antibody fragment according to claim 14, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 38, and the light chain variable region has any one of the amino acid sequences of SEQ ID NOs: 41 to 48.
  19. The antibody or its antigen-binding antibody fragment or bispecific antibody or its antigen-binding antibody fragment according to claim 14, wherein the heavy chain variable region has the amino acid sequence of SEQ ID NO: 39, and the light chain variable region has any one of the amino acid sequences of SEQ ID NOs: 41 to 48.
  20. An antibody or its antigen-binding antibody fragment or a bispecific antibody or its antigen-binding antibody fragment, having a ratio of at least 1.5:1, at least 2:1, at least 3:1, at least 4:1, at least 5:1, at least 6:1, at least 7:1, at least 8:1, at least 9:1, or at least 10:1 between the binding affinity to the HER2 protein at pH 6.0 in the tumor microenvironment and the binding affinity to the HER2 protein at pH 7.4 in the non-tumor microenvironment .

Description

This disclosure relates to anti-HER2 antibodies, anti-HER2 antibody fragments, anti-HER2 polyspecific antibodies, and immunoconjugates of such antibodies and antibody fragments, as well as the use of antibodies, antibody fragments, polyspecific antibodies, and immunoconjugates in diagnostic and therapeutic methods. Human epidermal growth factor receptor 2 (HER2) is a member of the epidermal growth factor receptor family that possesses tyrosine kinase activity. Receptor dimerization leads to autophosphorylation of tyrosine residues within the receptor's cytoplasmic domain, initiating various signaling pathways that contribute to cell proliferation and tumorigenesis. Further details on the role of HER2 in cancer can be found in numerous articles, such as "Human Epidermal Growth Factor Receptor 2 (HER2) in Cancers: Overexpression and Therapeutic Implications," Iqbal, Nida, and Iqbal, Naveed, Molecular Biology International, Volume 2014, Article ID 852748. Overexpression of the ERB-B2 gene (also known as the "HER2 gene") occurs in a significant proportion of breast cancers. HER2 protein overexpression is strongly associated with increased disease recurrence and poor prognosis. Drugs targeting the HER2 protein in breast cancer have shown significant positive effects in the treatment of HER2-positive breast cancer. HER2 protein overexpression also occurs in ovarian cancer, gastric cancer, lung adenocarcinoma, invasive forms of uterine cancer, gastric cancer, and salivary duct cancer. The HER2 protein is a target of the monoclonal antibody trastuzumab, Herceptin, which has been shown to be effective in cancers where the HER2 protein is overexpressed. Trastuzumab binding to the HER2 protein has been shown to increase p27, a protein that inhibits cell proliferation. Another monoclonal antibody, pertuzumab, is approved by the FDA for use in combination with trastuzumab. Pertuzumab inhibits the dimerization of HER2 and HER3 receptors. Other therapies targeting the HER2 protein are available or under development. There are at least four tests for HER2 overexpression: the ImmunoHistology test, which determines whether there is too much HER2 protein in cancer cells; the fluorescence In Situ hybridization test, which determines whether there is too many copies of the HER2 gene in cancer cells; the subtraction probe technology chromogenic In Situ hybridization test, which also determines whether there is too many copies of the HER2 gene in cancer cells; and the Information Dual In Situ hybridization test, which also determines whether there is too many copies of the HER2 gene in cancer cells. The present invention aims to provide anti-HER2 antibodies or antibody fragments with reduced or minimal side effects, particularly suitable for therapeutic and diagnostic use for the diagnosis and treatment of cancer. Some of these anti-HER2 antibodies or antibody fragments may have a higher binding affinity to HER2 proteins in the tumor microenvironment compared to HER2 proteins present in normal tissues. These anti-HER2 antibodies or antibody fragments typically have at least equivalent efficacy to known anti-HER2 antibodies or antibody fragments. In addition, these anti-HER2 antibodies or antibody fragments may exhibit reduced side effects compared to monoclonal anti-HER2 antibodies, including those known in the art due to their relatively low binding affinity to HER2 proteins in normal tissues. These advantages can provide more selective targeting of HER2 proteins, and as a result of the antibody's selectivity for HER2 proteins present in the tumor microenvironment, it may be possible to use higher doses of these anti-HER2 antibodies or antibody fragments, thereby enabling more effective therapeutic treatment without a corresponding increase in undesirable side effects. In one embodiment, the present invention provides an isolated polypeptide that specifically binds to the HER2 protein, wherein the polypeptide has a heavy chain variable region comprising three anti-HER2 complementarity determining regions, the region having sequences H1, H2, and H3. The H1 sequence is GFX 1 IKDTYIH (sequence number 1), The H2 sequence is X 2 IX 3 PTX 4 X 5 YX 6 X 7 YADSVKG (Sequence ID 2), The H3 sequence is WGGDGFYX 8 MDY (Sequence ID 3), In the formula, X1 is N or W, X2 is R or K, X3 is Y, K or D, X4 is N or A, X5 is G or K, X6 is T or D , X7 is R or E, and X8 is A or E, representing a heavy-chain variable region. A light chain variable region comprising three anti-HER2 complementarity determining regions having sequences L1, L2, and L3, The L1 sequence is RASQDVNTX 9 VA (sequence number 4), The L2 sequence is SASFLYS (sequence number 5), The L3 sequence is QQX 10 YTTPPPT (sequence number 6), The formula includes a light chain variable region where X9 is A or D and X10 is H, D, or E, As a premise, if X1 to X8 are N, R, Y, N, G, T, R, and A respectively, then X9 is not A and X10 is not H. The isolated polypeptide of the present invention has an