Trispecific proteins and methods of use
Provided herein are B cell maturation agent (BCMA) targeting trispecific proteins comprising a domain binding to CD3, a half-life extension domain, and a domain binding to BCMA. Also provided are pharmaceutical compositions thereof, as well as nucleic acids, recombinant expression vectors and host cells for making such BCMA targeting trispecific proteins. Also disclosed are methods of using the disclosed BCMA targeting trispecific proteins in the prevention, and/or treatment diseases, conditions and disorders.
1. A B cell maturation agent (BCMA) binding trispecific protein that comprises:
(a) a first domain (A) which is a single chain variable fragment (scFv) that specifically binds to a human CD3;
(b) a second domain (B) which is a single domain antibody that specifically binds to a human serum albumin protein; and
(c) a third domain (C) which is a single domain antibody that specifically binds to a human BCMA comprising the sequence of SEQ ID NO: 468,
wherein the third domain comprises complementarity determining regions CDR1, CDR2, and CDR3 and wherein
the CDR1 comprises the amino acid sequence of SEQ ID No. 76, the CDR2 comprises the amino acid sequence of SEQ ID No. 190, and the CDR3 comprises the amino acid sequence of SEQ ID No. 304.
2. The BCMA binding trispecific protein of claim 1 , wherein the first domain, the second domain, and the third domain are independently humanized.
3. The BCMA binding trispecific protein of claim 1 , wherein the third domain comprises the following formula:
f1-r1-f2-r2-f3-r3-f4
wherein, r1 is the CDR1; r2 is the CDR2; and r3 is the CDR3; and wherein f1, f2, f3 and f4 are framework residues selected so that said protein is from about 80% to about 99% identical to the amino acid sequence set forth in SEQ ID NO: 346 or SEQ ID NO: 598.
4. The BCMA binding trispecific protein of claim 3 , wherein f1 comprises SEQ ID NO: 461 or 462, f2 comprises SEQ ID NO: 463, f3 comprises SEQ ID NO: 464 or 465, and f4 comprises SEQ ID NO: 466 or 467.
5. The BCMA binding trispecific protein of claim 1 , wherein the third domain comprises a4 the amino sequence.
6. The BCMA binding trispecific protein of claim 1 , wherein said protein has an elimination half-time of at least 12 hours, at least 20 hours, at least 25 hours, at least 30 hours, at least 35 hours, at least 40 hours, at least 45 hours, at least 50 hours, or at least 100 hours, when administered to a subject, wherein the subject is a human.
7. The BCMA binding trispecific protein of claim 2 , wherein the third domain binds to an extracellular domain of BCMA.
8. The BCMA binding trispecific protein of claim 1 , wherein the trispecific protein comprises the sequence of SEQ ID NO: 520.
9. The BCMA binding trispecific protein of claim 1 , that further comprises linkers L1 and L2, wherein the domains of the protein are linked in the order H2N-(A)-L1-(C)-L2-(B)-COOH, H2N-(B)-L1-(A)-L2-(C)-COOH, H2N-(C)-L1-(B)-L2-(A)-COOH, H2N-(C)-L1-(A)-L2-(B)-COOH, H2N-(A)-L1-(B)-L2-(C)-COOH or H2N-(B)-L1-(C)-L2-(A)-COOH.
10. The BCMA binding trispecific protein of claim wherein the domains are linked in the order H 2 N-(C)-L1-(B)-L2-(A)-COOH.
11. The BCMA binding trispecific protein of claim 10 , wherein the linkers L1 and L2 are each independently selected from a group consisting of (GS) n (SEQ ID NO: 472), (GGS) n (SEQ ID NO: 473), (GGGS) n (SEQ ID NO: 474), (GGSG) n (SEQ ID NO: 475), (GGSGG) n (SEQ ID NO: 476), (GGGGS) n (SEQ ID NO: 477), (GGGGG) n (SEQ ID NO: 478) and (GGG) n (SEQ ID NO: 479) wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.
12. A method for the treatment or amelioration of a tumorous disease associated with BCMA in a subject having the tumorous disease, comprising administering to the subject a pharmaceutical composition that comprises the BCMA binding trispecific protein of claim 1 .
13. A method for the treatment or amelioration of a tumorous disease associated with BCMA in a subject having the tumorous disease, comprising administering to the subject a pharmaceutical composition that comprises the BCMA binding trispecific protein of claim 8 .