Multispecific antibodies, compositions comprising the same, and vectors and uses thereof
The present disclosure provides multispecific antibodies having increased in vivo sustainability, the multispecific antibodies comprising one or more bioactive effector moieties linked to either or both of an N-terminal and a C-terminal of an antigen binding fragment Fab that binds to human serum albumin.
1. A multispecific antibody comprising a structural formula of:
wherein the antigen binding fragment is a serum albumin Fab comprising
a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74), a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76), and
a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91), a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93);
wherein R 1 and R 2 are bioactive effector moieties linked to an N-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab, wherein each of the R 1 and R 2 is an anti-CD40L hu5c8 scFv comprising an amino acid sequence of SEQ ID NO:48;
wherein R 3 and R 4 are bioactive effector moieties linked to a C-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab;
wherein m is 0; and
wherein n is 0.
2. The multispecific antibody of claim 1 , wherein each of R 1 and R 2 is linked to the Fab by one or more linkers.
3. The multispecific antibody of claim 1 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 99.
4. The multispecific antibody of claim 1 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO: 99.
5. The multispecific antibody of claim 1 , wherein the Fab comprises a light chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 105.
6. The multispecific antibody of claim 1 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 99, and a light chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 105.
7. The multispecific antibody of claim 1 , wherein the Fab comprises a heavy chain domain comprising an amino acid sequence of SEQ ID NO:45 (V H -C H1 domain) and a light chain domain comprising an amino acid sequence of SEQ ID NO:46 (V L -C L domain).
8. The multispecific antibody of claim 2 , wherein each linker comprises 1 to 20 amino acids.
9. The multispecific antibody of claim 2 , wherein each linker comprises an amino acid sequence having at least 90% identity to SEQ ID NO:3 or SEQ ID NO:4.
10. The multispecific antibody of claim 1 , wherein the multispecific antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:41, and a light chain comprising an amino acid sequence of SEQ ID NO:42.
11. A multispecific antibody comprising a structural formula of:
wherein the antigen binding fragment is a serum albumin Fab comprising
a heavy chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of AYSMN (SEQ ID NO:74), a heavy chain CDR2 comprising the amino acid sequence of SISSSGRYIHYADSVKG (SEQ ID NO:75), and a heavy chain CDR3 comprising the amino acid sequence of ETVMAGKALDY (SEQ ID NO:76), and
a light chain complementarity determining domain 1 (CDR1) comprising the amino acid sequence of RASQSVGSNLA (SEQ ID NO:91), a light chain CDR2 comprising the amino acid sequence of GASTGAT (SEQ ID NO:92), and a light chain CDR3 comprising the amino acid sequence of QQYYSFLAKT (SEQ ID NO:93);
wherein R 1 and R 2 are bioactive effector moieties linked to an N-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab, wherein each of the R 1 and R 2 is an anti-CD40L hu5c8 scFv comprising an amino acid sequence of SEQ ID NO:48;
wherein R 3 and R 4 are bioactive effector moieties linked to a C-terminus of the Fab, each linked to a heavy chain variable domain or a light chain variable domain of the Fab;
wherein m is 0;
wherein n is 0; and
wherein each of R 1 and R 2 is linked to the Fab by one or more linkers, wherein each linker comprises an amino acid sequence of SEQ ID NO:3 or SEQ ID NO:4.
12. The multispecific antibody of claim 11 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 99.
13. The multispecific antibody of claim 11 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence of SEQ ID NO: 99.
14. The multispecific antibody of claim 11 , wherein the Fab comprises a light chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 105.
15. The multispecific antibody of claim 11 , wherein the Fab comprises a heavy chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 99, and a light chain variable domain comprising an amino acid sequence having at least 98% identity to SEQ ID NO: 105.
16. The multispecific antibody of claim 11 , wherein the Fab comprises a heavy chain domain comprising an amino acid sequence of SEQ ID NO:45 (V H -C H1 domain) and a light chain domain comprising an amino acid sequence of SEQ ID NO:46 (V L -C L domain).
17. The multispecific antibody of claim 11 , wherein the multispecific antibody comprises a heavy chain comprising an amino acid sequence of SEQ ID NO:41, and a light chain comprising an amino acid sequence of SEQ ID NO:42.
18. A pharmaceutical composition comprising the multispecific antibody of claim 1 and a pharmaceutically acceptable excipient.
19. A pharmaceutical composition comprising the multispecific antibody of claim 11 and a pharmaceutically acceptable excipient.