(19) TZZ ¥¥ _T (11) EP 2 673 362 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12N 15/113 (2010.01) A61K 31/713 (2006.01) 07.06.2017 Bulletin 2017/23 A61P 35/04 (2006.01) A61K 39/395 (2006.01) C07K 16/30 (2006.01) (21) Application number: 12706220.6 (86) International application number: (22) Date of filing: 09.02.2012 PCT/EP2012/052238 (87) International publication number: WO 2012/107531 (16.08.2012 Gazette 2012/33) (54) PLEXIN-B1 ANTAGONISTS AND USES THEREOF PLEXIN-B1 ANTAGONISTEN UND VERWENDUNGEN DAVON ANTAGONISTES DE PLEXINE B1 ET UTILISATIONS DE CEUX-CI (84) Designated Contracting States: • DEAGLIO SILVIA ET AL: "CD38 and CD100 lead AL AT BE BG CH CY CZ DE DK EE ES FI FR GB a network of surface receptors relaying positive GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO signals for B-CLL growth and survival", BLOOD, PL PT RO RS SE SI SK SM TR AMERICAN SOCIETY OFHEMATOLOGY, US, vol. 105, no. 8, 1 April 2005 (2005-04-01) , pages (30) Priority: 09.02.2011 EP 11153784 3042-3050, XP002405817, ISSN: 0006-4971, DOI: 10.1182/BLOOD-2004-10-3873 (43) Date of publication of application: • SWIERCZ JAKUB M ET AL: "Semaphorin 4D 18.12.2013 Bulletin 2013/51 signaling requires the recruitment of phospholipase C gamma into the plexin-B1 (73) Proprietors: receptor complex.", MOLECULAR AND • Ruprecht-Karls-Universität Heidelberg CELLULAR BIOLOGY, vol. 29, no. 23, December 69117 Heidelberg (DE) 2009 (2009-12), pages 6321-6334, XP002640895, • Max-Planck-Gesellschaft zur Förderung ISSN: 1098-5549 cited in the application der Wissenschaften e.V. • SWIERCZ J M ET AL: "ErbB-2 and met 80539 München (DE) reciprocally regulate cellular signaling via plexin-B1", JOURNAL OF BIOLOGICAL (72) Inventors: CHEMISTRY, vol. 283, no. 4, 25 January 2008 • OFFERMANNS, Stefan (2008-01-25), pages 1893-1901, XP002640896, 61231 Bad Nauheim (DE) AMERICAN SOCIETY FOR BIOCHEMISTRY AND • SWIERCZ, Jakub MOLECULAR BIOLOGY INC. US DOI: 61231 Bad Nauheim (DE) 10.1074/JBC.M706822200 • WORZFELD, Thomas • YE SHUANGMEI ET AL: "Plexin-B1 silencing 61231 Bad Nauheim (DE) inhibits ovarian cancer cell migration and invasion.", BMC CANCER, vol. 10, 611, 2010, (74) Representative: Herzog, Fiesser & Partner pages 1-11, XP002640894, ISSN: 1471-2407 Patentanwälte PartG mbB • BERT J. C. JANSSEN ET AL: "Structural basis of Dudenstrasse 46 semaphorin-plexin signalling", NATURE, vol. 68167 Mannheim (DE) 467, no. 7319, 28 October 2010 (2010-10-28), pages 1118-1122, XP55025500, ISSN: 0028-0836, (56) References cited: DOI: 10.1038/nature09468 WO-A2-01/14420 WO-A2-03/031930 WO-A2-2006/110593 WO-A2-2010/129917 Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 673 362 B1 Printed by Jouve, 75001 PARIS (FR) (Cont. next page) EP 2 673 362 B1 • MUTSUMI HIROTANI ET AL: "Interaction of • THOMAS WORZFELD ET AL: "Semaphorins and plexin-B1 with PDZ domain-containing Rho plexins as therapeutic targets", NATURE guanine nucleotide exchange factors", REVIEWS DRUG DISCOVERY, vol. 13, no. 8, 1 BIOCHEMICAL AND BIOPHYSICAL RESEARCH August 2014 (2014-08-01) , pages 603-621, COMMUNICATIONS, vol. 297, no. 1, 1 September XP055167541, ISSN: 1474-1776, DOI: 2002 (2002-09-01), pages 32-37, XP55025568, 10.1038/nrd4337 ISSN: 0006-291X, DOI: • I. Chabbert-De Ponnat ET AL: "Soluble CD100 10.1016/S0006-291X(02)02122-8 functions on human monocytes and immature • WORZFELD T ET AL: "HER2/ErbB2 signals dendritic cells require plexin C1 and plexin B1, through Plexin-B1 to promote breast cancer respectively", INTERNATIONAL IMMUNOLOGY., metastasis", NAUNYN-SCHMIEDEBERG’S vol. 17, no. 4, 3 March 2005 (2005-03-03), pages ARCHIVES OF PHARMACOLOGY, vol. 383, no. 439-447, XP055320934, GB ISSN: 0953-8178, DOI: Suppl. 1, March 2011 (2011-03), page 3, 10.1093/intimm/dxh224 XP002674682, & 77TH ANNUAL MEETING ON • Kazunori Yukawa ET AL: "Sema4A induces cell GERMAN-SOCIETY-FOR-EXPERIMENTAL-AND- morphological changes through B-type CLINIC AL-P plexin-mediatedsignaling", International Journal HARMACOLOGY-AND-TOXICOLOGY; of Molecular Medicine, 1 February 2010 FRANKFURT, GERMANY; MARCH 30 -APRIL 01, (2010-02-01), pages 225-230, XP055320937, DOI: 2011 10.3892/ijmm_00000334 Retrieved from the • SWIERCZ JAKUB M ET AL: Internet: "Plexin-B1/RhoGEF-mediated RhoA activation URL:https://www.spandidos-publications.com involves the receptor tyrosine kinase ErbB-2.", /ijmm/25/2/225/download [retrieved on THE JOURNAL OF CELL BIOLOGY, vol. 165, no. 2015-08-11] 6, 21 June 2004 (2004-06-21) , pages 869-880, • S. Artigiani: "Functional Regulation of XP002640897, ISSN: 0021-9525 cited in the Semaphorin Receptors by Proprotein application Convertases", Journal of Biological Chemistry, • SUN QIANGMING ET AL: "Hypoxia-inducible vol. 278, no. 12, 17 January 2003 (2003-01-17), factor-1-mediated regulation of semaphorin 4D pages 10094-10101, XP055320938, US ISSN: affects tumor growth and vascularity.", THE 0021-9258, DOI: 10.1074/jbc.M210156200 JOURNAL OF BIOLOGICAL CHEMISTRY, vol. • SREEHARSHA GURRAPU ET AL: 284, no. 46, 13 November 2009 (2009-11-13), "Transmembrane semaphorins: Multimodal pages 32066-32074, XP002640898, ISSN: signaling cues in development and cancer", 1083-351X CELL ADHESION AND MIGRATION, 13 June 2016 • THOMAS WORZFELD ET AL: "ErbB-2 signals (2016-06-13), pages 1-17, XP055320941, US ISSN: through Plexin-B1 to promote breast cancer 1933-6918, DOI: 10.1080/19336918.2016.1197479 metastasis", J CLIN INVEST, vol. 122, no. 4, 1 • BATTISTINI CHIARA ET AL: "Transmembrane March 2012 (2012-03-01) , pages 1296-1305, semaphorins, forward and reverse signaling: XP55025554, DOI: 10.1172/JCI60568 have a look both ways", CMLS CELLULAR AND • Kumanogoh ET AL: "Identification of CD72 as a MOLECULAR LIFE SCIENCES, BIRKHAUSER lymphocyte receptor for the class IV semaphorin VERLAG, HEIDELBERG, DE, vol. 73, no. 8, 21 CD100: a novel mechanism for regulating B cell January 2016 (2016-01-21), pages 1609-1622, signaling.", Immunity, vol. 13, no. 5, 1 November XP035801849, ISSN: 1420-682X, DOI: 2000 (2000-11-01), pages 621-631, XP055039972, 10.1007/S00018-016-2137-X [retrieved on ISSN: 1074-7613, DOI: 2016-01-21] 10.1016/S1074-7613(00)00062-5 • DEBORAH A. WITHERDEN ET AL: "The CD100 Remarks: Receptor Interacts with Its Plexin B2 Ligand to Thefile contains technical information submitted after Regulate Epidermal [gamma][delta] T Cell the application was filed and not included in this Function", IMMUNITY., vol. 37, no. 2, 1 August specification 2012 (2012-08-01) , pages 314-325, XP055306038, US ISSN: 1074-7613, DOI: 10.1016/j.immuni.2012.05.026 • KENTA MASUDA ET AL: "Sema4D stimulates axonal outgrowth of embryonic DRG sensory neurones", GENES TO CELLS, WILEY-BLACKWELL PUBLISHING LTD, GB, vol. 9, no. 9, 1 September 2004 (2004-09-01), pages 821-829, XP008144069, ISSN: 1356-9597, DOI: 10.1111/J.1365-2443.2004.00766.X [retrieved on 2004-08-25] 2 EP 2 673 362 B1 Description [0001] The present invention concerns the field of cancer therapy. In particular, it relates to an antagonist of plexin- B1 which prevents the interaction of the plexin-B1 with ErbB-2 for use as a medicament, wherein said antagonist is an 5 antibody specifically binding to the extracellular domain of the plexin-B1 polypeptide and inhibiting binding of said plexin- B1 polypeptide to ErbB-2, but not inhibiting the binding of the ligand Sema4D to its receptor plexin-B1 polypeptide. The invention pertains also to an antagonist of plexin-B1 which prevents the binding of the ligand Sema4D to its receptor plexin-B1 polypeptide for use as a medicament, wherein said antagonist is an antibody specifically binding to the extra- cellular domain of the plexin-B1 polypeptide and inhibiting the binding of the ligand Sema4D to its receptor plexin-B1 10 polypeptide, wherein the antibody i) shows specific binding in Western blot and immunoblot at a concentration of 10 microgram/ml; ii) is able to precipitate plexin-B1 overexpressed in HEK cells and native plexin-B1 in MCF-7 and BT-474 cells; iii) is able to co-immunoprecipitate plexin-B1/ErbB-2 complex in MCF-7, BT-474 and SK-OV-3 cells; 15 iv) does not block basal RhoA activity in BT-474 and SK-OV-3 cells, but blocks Sema4D-dependent RhoA activation in MCF-7 cells, due to the competitive inhibition of binding of the ligand Sema4D to the receptor plexin-B1; v) blocks R-Ras deactivation via plexin-B1 after stimulation with Sema4D in MCF-7 cells; and vi) binds specifically to plexin-B1, and exhibits no cross-reactivity to other plexin-family members. 20 [0002] Described is a method for identifying an antagonist which prevents the interaction of a B-type plexin with ErbB- 2. Finally, a polynucleotide encoding a B-type plexin polypeptide which lacks a functional intracellular domain and the said polypeptide is described. [0003] Breast cancer is the most common primary malignancy in women. About 30% of all breast cancers overexpress the receptor tyrosine kinase ErbB-2 (Slamon 1989, Science 244:707-712). These tumors are characterized by aggressive 25 behavior and poor prognosis. A plethora of evidence including transgenic mice which overexpress ErbB-2 in their mam- mary glands and subsequently develop breast cancer directly implicates ErbB-2 signalling in mammary oncogenesis (Muller 1988, Cell 54:105-15; Guy 1992, Proc Natl Acad Sci U S A 89:10578-82).