US 11,952,418 B2
Biparatopic polypeptides antagonizing Wnt signaling in tumor cells
Vittoria Zinzalla, Vienna (AT); Klaus-Peter Kuenkele, Perchtoldsdorf (AT); Marie-Ange Buyse, Merelbeke (BE); Karen Cromie, Zwijnaarde (BE); Stephanie Staelens, Zwijnaarde (BE); and Beatrijs Strubbe, Zwijnaarde (BE)
Assigned to Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed by Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed on Feb. 12, 2020, as Appl. No. 16/788,329.
Application 16/788,329 is a continuation of application No. 15/367,323, filed on Dec. 2, 2016, granted, now 10,597,449.
Claims priority of application No. 15197999 (EP), filed on Dec. 4, 2015.
Prior Publication US 2020/0199222 A1, Jun. 25, 2020
Int. Cl. A61K 39/00 (2006.01); C07K 16/18 (2006.01); C07K 16/28 (2006.01)
CPC C07K 16/28 (2013.01) [C07K 16/18 (2013.01); A61K 2039/505 (2013.01); A61K 2039/545 (2013.01); C07K 2317/22 (2013.01); C07K 2317/24 (2013.01); C07K 2317/31 (2013.01); C07K 2317/33 (2013.01); C07K 2317/40 (2013.01); C07K 2317/565 (2013.01); C07K 2317/567 (2013.01); C07K 2317/569 (2013.01); C07K 2317/62 (2013.01); C07K 2317/70 (2013.01); C07K 2317/73 (2013.01); C07K 2317/76 (2013.01); C07K 2317/92 (2013.01); C07K 2317/94 (2013.01); C07K 2319/31 (2013.01)] 3 Claims
 
1. A method of treatment of tumors expressing human LRP5 and/or human LRP6 by inhibition of the Wnt-1 and/or Wnt-3a signaling pathways in a cancer patient comprising administering to said patient a cross-reactive biparatopic polypeptide which specifically binds to an epitope of the human low-density receptor-like protein 5 (LRP5) as well as an epitope of the human low-density receptor-like protein 6 (LRP6) in an amount effective to reduce said patient's tumor burden and/or tumor size wherein said cross-reactive biparatopic polypeptide comprises:
(a) a first immunoglobulin single variable domain, wherein said immunoglobulin single variable domain is a humanized VHH domain capable of inhibiting the Wnt1 signaling pathway comprising the CDR sequences:
 
 (SEQ ID NO: 4)
 
 CDR1: SYAMG
 
 
 
 (SEQ ID NO: 5)
 
 CDR2: AIRRSGRRTYYADSVKG
 
 and
 
 
 
 (SEQ ID NO: 6)
 
 CDR3: ARRVRSSTRYNTGTWWWEY;
and
(b) a second immunoglobulin single variable domain wherein said second immunoglobulin single variable domain is a humanized VHH domain capable of inhibiting the Wnt3a signaling pathway comprising the following CDR sequences:
 
 (SEQ ID NO: 13)
 
 CDR1: SYAMG
 
 
 
 (SEQ ID NO: 14)
 
 CDR2: AISWRSGSTYYADSVKG
 
 and
 
 
 
 (SEQ ID NO: 15)
 
 CDR3: DPRGYGVAYVSAYYEY.
wherein said first and said second immunoglobulin single variable domains are covalently linked by a linker peptide, wherein said linker peptide comprises a third immunoglobulin single variable domain which binds to albumin comprising SEQ ID NO:24.